ExperimentELISA
ExperimentELISA[Samples]⟹Protocol
creates a Protocol to run Enzyme-Linked Immunosorbent Assay (ELISA) experiment on the provided Samples for the detection and quantification of certain analytes.
Enzyme-Linked Immunosorbent Assay (ELISA) uses antigen-antibody interaction to quantitatively measure the concentration of an antigen in samples or compare antibody binding affinity. There are many forms of ELISA, such as direct ELISA, indirect ELISA, sandwich ELISA, competitive ELISA, and fast ELISA that apply antigens and antibodies in varied sequences. During an ELISA experiment, protein standards and samples are loaded onto ELISA plates and the target antigen is bound by antibodies. Antigen concentration is revealed by an enzyme, which converts its substrate into a colored product. Results are measured using colorimetric detection, and antigen concentrations are calculated using the standard curve generated from protein standards.
Experimental Principles
Figure 1.1: Overview of ELISA methods. In a DirectELISA experiment, the Sample is coated on the ELISAPlate, and then a primary antibody is used to detect the target antigen. In a DirectSandwichELISA experiment, a capture antibody is coated onto the ELISAPlate to pull down the target antigen from the sample. Then primary antibody is used to detect the target antigen. In a DirectCompetitiveELISA experiment, a reference antigen is used to coat the ELISAPlate. Samples are incubated with the primary antibody. Then, when the Sample-Antibody-Complex solution is loaded on the ELISAPlate, the remaining free primary antibody binds to the reference antigen. In a FastELISA experiment, a coating antibody against a tag is coated to the ELISAPlate. A capture antibody containing this small tag and a primary antibody for antigen detection is incubated with the sample to form a CaptureAntibody-TargetAntigen-PrimaryAntibody complex. Then this complex is pulled down by the coating antibody to the surface of the plate. Compared with the Direct methods where the primary antibody is conjugated with an enzyme (such as Horse Radish Peroxidase/HRP or Alkaline Phosphatas/AP) for detection, in Indirect methods a secondary antibody is, instead, conjugated with this enzyme. During Indirect methods, an additional step of SecondaryAntibody incubation is added to the corresponding Direct methods.
Figure 1.2: DirectELISA process: 1. If required, antibodies are diluted. 2. Sample, Spike, Standard, Blank and any dilutions are assembled in 96-well 2ml deep well plate. 3. Assembled samples from the previous step are transferred to the ELISA plate and incubated to coat the plate. 4. Blocking buffer is added to the ELISA plate to prevent non-specific binding. 5. Primary antibody is added to the plate for immunosorbence. 5. SubstrateSolution and StopSolution (if required) are added to the plate for colorimetric detection.
Figure 1.3: IndirectELISA process: 1. If required, antibodies are diluted. 2. Sample, Spike, Standard, Blank and any dilutions are assembled in 96-well 2ml deep well plate. 3. Assembled samples from the previous step are transferred to the ELISA plate and incubated to coat the plate. 4. Blocking buffer is added to the ELISA plate to prevent non-specific binding. 5. Primary antibody and secondary antibody are sequentially added to the plate for immunosorbence. 5. SubstrateSolution and StopSolution (if required) are added to the plate for colorimetric detection.
Figure 1.4: DirectSandwichELISA process: 1. If required, antibodies are diluted. 2. Capture antibody is added to the ELISA plate for coating. 3. Sample, Spike, Standard, Blank and any dilutions are assembled in 96-well 2ml deep well plate. 4. Blocking buffer is added to the ELISA plate to prevent non-specific binding. 5. Assembled samples and primary antibody are added to the plate sequentially for immunosorbence. 6. SubstrateSolution and StopSolution (if required) are added to the plate for colorimetric detection.
Figure 1.5: IndirectSandwichELISA process: 1. If required, antibodies are diluted. 2. Capture antibody is added to the ELISA plate for coating. 3. Sample, Spike, Standard, Blank and any dilutions are assembled in 96-well 2ml deep well plate. 4. Blocking buffer is added to the ELISA plate to prevent non-specific binding. 5. Assembled samples, primary antibody, and secondary antibody are added to the plate sequentially for immunosorbence. 6. SubstrateSolution and StopSolution (if required) are added to the plate for colorimetric detection.
Figure 1.6: DirectCompetitiveELISA process: 1. If required, antibodies and reference antigen are diluted. 2. Reference antigen is added to the ELISA plate for coating. 3. Sample, Spike, Standard, Blank and any dilutions are assembled in 96-well 2ml deep well plate and primary antibody is mixed with each sample and incubated (if required). 4. Blocking buffer is added to the ELISA plate to prevent non-specific binding. 5. The Sample-antibody mixture from step 3 is added to the ELISA plate for immunosorbence. 6. SubstrateSolution and StopSolution (if required) are added to the plate for colorimetric detection.
Figure 1.7: IndirectCompetitiveELISA process: 1. If required, antibodies and reference antigen are diluted. 2. Reference antigen is added to the ELISA plate for coating. 3. Sample, Spike, Standard, Blank and any dilutions are assembled in 96-well 2ml deep well plate and primary antibody is mixed with each sample and incubated (if required). 4. Blocking buffer is added to the ELISA plate to prevent non-specific binding. 5. The Sample-antibody mixture from step 3 is added to the ELISA plate, and then the secondary antibody is added, for immunosorbence. 6. SubstrateSolution and StopSolution (if required) are added to the plate for colorimetric detection.
Figure 1.8: FastELISA process: 1. If required, antibodies are diluted. 2. Coating antibody is added to the ELISA plate for coating. 3. Sample, Spike, Standard, Blank and any dilutions are assembled in 96-well 2ml deep well plate. Then primary antibody and capture antibody are mixed with each sample and incubated (if required). 4. Blocking buffer is added to the ELISA plate to prevent non-specific binding. 5. The Sample-antibody mixture from step 3 is added to the ELISA plate for immunosorbence. 6. SubstrateSolution and StopSolution (if required) are added to the plate for colorimetric detection.
Experiment Options
General
Method
Defines the type of ELISA experiment to be performed. Types include DirectELISA, IndirectELISA, DirectSandwichELISA, IndirectSandwichELISA, DirectCompetitiveELISA, IndirectCompetitiveELISA, and FastELISA. Compared with the Direct methods where the primary antibody is conjugated with an enzyme (such as Horse Radish Peroxidase/HRP or Alkaline Phosphatas/AP) for detection, in all Indirect methods a secondary antibody is, instead, conjugated with this enzyme. In Indirect methods, an additional step of SecondaryAntibody incubation is added to the corresponding Direct methods. In a DirectELISA experiment, the Sample is coated on the ELISAPlate, and then a primary antibody is used to detect the target antigen. In a DirectSandwichELISA experiment, a capture antibody is coated onto the ELISAPlate to pull down the target antigen from the sample. Then a primary antibody is used to detect the target antigen. In a DirectCompetitiveELISA experiment, a reference antigen is used to coat the ELISAPlate. Samples are incubated with the primary antibody. Then, when the Sample-Antibody-Complex solution is loaded on the ELISAPlate, the remaining free primary antibody binds to the reference antigen. In a FastELISA experiment, a coating antibody against a tag is coated to the ELISAPlate. A capture antibody containing this tag and a primary antibody for antigen detection is incubated with the sample to form a CaptureAntibody-TargetAntigen-PrimaryAntibody complex. Then this complex is pulled down by the coating antibody to the surface of the plate (See Figure 1.1).
Pattern Description: DirectELISA, IndirectELISA, DirectSandwichELISA, IndirectSandwichELISA, DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA.
TargetAntigen
The Analyte molecule (e.g., peptide, protein, and hormone) detected and quantified in the samples by Antibodies in the ELISA experiment. This option is used to automatically set sample Antibodies and the corresponding experiment conditions of Standards and Blanks.
NumberOfReplicates
The number of times an ELISA assay will be repeated in parallel wells. Samples, Standards, or Blanks will be loaded this number of times in the plates. Replications are conducted at the same time, and if possible, in the same ELISAPlate. If set to Null, each Sample, Standard, or Blank will be assayed once.
Default Calculation: Automatically set to 2 unless method is DirectELISA or IndirectELISA and Coating is False, in which case it is set to Null.
Pattern Description: Greater than or equal to 2 and less than or equal to 192 in increments of 1 or Null.
WashingBuffer
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample.
Sample Assembly
Spike
The sample with a known concentration of analyte. Spike is to be mixed with the input sample. The purpose of spiking is to perform a spike-and-recovery assessment to determine whether the ELISA can accurately test the concentration of analyte within the context of the sample. If the recovery observed for the spiked sample is identical to the analyte prepared in standard diluent, the sample is considered not to interfere with the assay. For example, if molecules in the sample inhibits the binding between the antibody and TargetAntigen, the results from the ELISA becomes inaccurate. In a Spike-and-Recovery experiment,an aliquot of a sample is mixed with Spike. ELISA is performed on the Sample alone (also called Neat Sample) and the Spiked Sample at the same time, where the Spike concentration in the sample can be measured. This measured Spike concentration can be then compared with the known Spike concentration. Typically a 20% difference between measured Spike concentration and the known Spike concentration is acceptable. Spiked sample can be further diluted to perform linearity-of-dilution assessment. The plate used for sample spiking and dilution will be discarded after the experiment.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
SpikeDilutionFactor
The dilution ratio by which the Spike is mixed with the input sample before further dilution is performed.
SpikeStorageCondition
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
SampleSerialDilutionCurve
The collection of serial dilutions that will be performed on each sample: if spiking was performed, the dilution will be performed on the spiked sample; otherwise, the dilutions will be performed on the sample. This Linearity-of-Dilution step assesses whether the ELISA experiment can reliably measure TargetAntigen concentration at different concentration ranges. For example, a sample with TargetAntigen concentration of 100ng/ul, if diluted 1:2, 1:4, and 1:8, should yield ELISA measurements of 50ng/ul, 25ng/ul, and 12.5ng/ul (or should follow a concentration ratio of 4:2:1). Typically a 20% difference between the measured and expected concentrations is considered acceptable. For Serial Dilution Volumes, the Transfer Volume is taken out of the sample and added to a second well with the Diluent Volume of the Diluent. It is mixed, then the Transfer Volume is taken out of that well to be added to a third well. This is repeated to make Number Of Dilutions diluted samples. For example, if a 100 ug/ ml sample with a Transfer Volume of 20 Microliters, a Diluent Volume of 60 Microliters and a Number of Dilutions of 3 is used, it will create a DilutionCurve of 25 ug/ ml, 6.25 ug/ ml, and 1.5625 ug/ ml with each dilution having a volume of 60 Microliters. For Serial Dilution Factors, the sample will be diluted by the dilution factor at each transfer step. IMPORTANT: because the dilution curve does not intrinsically include the original sample, in the case of sample dilution the first diluting factor should be 1 or Diluent Volume should be 0 Microliter to include the original sample. For example, if a Spike-and-Recovery assay is to be performed, the first dilution factor must be 1 in order to include the original spiked sample. During experiment, an 5% extra volume than specified is going to be prepared in order to offset pipetting errors. Therefore, the total volume for each well specified in this option should be equal or greater than the volume needed for the experiment. Because the dilution of each assay (well) is prepared independently, the Number of Replications should not be considered when determining the Spike Volume. The plate used for sample dilutions will be discarded after the experiment. Note: if spike or antibodies are to be mixed with samples, their volumes are counted towards diluent volume.
Figure 3.1: Use the SerialDilutionCuve option to create a collection of serial dilutions that will be performed on samples.
Programmatic Pattern: (({RangeP[10*Microliter, 1500*Milliliter], {RangeP[0, 1], GreaterP[0, 1]} | {RangeP[0, 1]..}} | {RangeP[0*Microliter, 1000*Microliter], RangeP[0*Microliter, 1000*Microliter], GreaterP[0, 1]}) | Automatic) | Null
SampleDilutionCurve
The collection of dilutions that will be performed on each sample: the dilutions will be performed on the sample. Spiked samples should only be further diluted with serial dilution. This Linearity-of-Dilution step assesses whether the ELISA experiment can reliably measure TargetAntigen concentration at different concentration ranges. For example, a sample with TargetAntigen concentration of 100ng/ul, if diluted 1:2, 1:4, and 1:8, should yield ELISA measurements of 50ng/ul, 25ng/ul, and 12.5ng/ul (or should follow a concentration ratio of 8:4:2:1). Typically a 20% difference between the measured and expected concentrations is considered acceptable. Linearity-of-Dilution experiment can be done with Spiked or non-Spiked samples. For Fixed Dilution Volume Dilution Curves, the Spike Amount is the volume of the sample that will be mixed the Diluent Volume of the Diluent to create a desired concentration. The Assay Volume is the TOTAL volume of the sample that will be created after being diluted by the Dilution Factor. IMPORTANT: because the dilution curve does not intrinsically include the original sample, in the case of sample dilution the first diluting factor must be 1, or Diluent Volume should be 0 to include the original sample. During experiment, an 5% extra volume than specified is going to be prepared in order to offset pipetting errors. Therefore, the total volume for each well specified in this option should be equal or greater than the volume needed for the experiment. Because the dilution of each assay (well) is prepared independently, the Number of Replications should not be considered when determining the Spike Volume. The plate used for sample dilutions will be discarded after the experiment. Note: if spike or antibodies are to be mixed with samples, their volumes are counted towards diluent volume.
Figure 3.2: Use the DilutionCuve option to create a collection of dilutions that will be performed on samples.
Default Calculation: The option is automatically set Null if or a SampleSerialDilutionCurve is specified.
Programmatic Pattern: (({{RangeP[10*Microliter, 1500*Microliter], RangeP[0, 1]}..} | {{RangeP[0*Microliter, 1000*Microliter], RangeP[0*Microliter, 1000*Microliter]}..}) | Automatic) | Null
SampleDiluent
Default Calculation: If SampleDilutionCurve and SampleSerialDilutionCurve are not both Null, in the case when Method is set to DirectELISA and IndirectELISA, the option is automatically set to Model[Sample, StockSolution, "1x Carbonate-Bicarbonate Buffer pH10"]; in all other cases, the option is automatically set to Model[Sample,"ELISA Blocker Blocking Buffer"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
Antibody Antigen Preparation
CoatingAntibody
Default Calculation: If Method is FastELISA, automatically set to an antibody against a tag which is conjugated to a the CaptureAntibody.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
CoatingAntibodyDilutionFactor
The dilution ratio of CoatingAntibody. CoatingAntibody is diluted with CoatingAntibodyDiluent. Either CoatingAntibodyDilutionFactor or CoatingAntibodyVolume should be provided but not both.
CoatingAntibodyVolume
The volume of undiluted CoatingAntibody added into the corresponding well of the assay plate. CoatingAntibody is diluted with CoatingAntibodyDiluent. CoatingAntibodyVolume is used as an alternative to CoatingAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
CoatingAntibodyDiluent
Default Calculation: If Method is set to FastELISA, the option is automatically set to Model[Sample, StockSolution, "1x Carbonate-Bicarbonate Buffer pH10"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
CoatingAntibodyStorageCondition
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
CaptureAntibody
The sample containing the antibody that is used to pull down the antigen from sample solution to the surface of the assay plate wells in DirectSandwichELISA, IndirectSandwichELISA, and FastELISA.
Default Calculation: If Method is FastELISA, automatically set to an antibody containing an affinity tag and against the TargetAntigen. If Method is DirectSandwichELISA or IndirectSandwichELISA, automatically set to an un-tagged antibody against TargetAntigen.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
CaptureAntibodyDilutionFactor
The dilution ratio of CaptureAntibody. For DirectSandwichELISA and IndirectSandwichELISA, CaptureAntibody is diluted with CaptureAntibodyDiluent. For FastELISA, CaptureAntibody is diluted in the corresponding sample. Either CaptureAntibodyDilutionFactor or CaptureAntibodyVolume should be provided but not both.
Default Calculation: For DirectSandwichELISA and IndirectSandwichELISA, automatically set to 0.001 (1:1,000). For FastELISA, automatically set to 0.01 (1:100).
CaptureAntibodyVolume
The volume of undiluted CaptureAntibody added into the corresponding well of the assay plate. CaptureAntibodyVolume is used as an alternative to CaptureAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
CaptureAntibodyDiluent
The buffer used to dilute CaptureAntibodies. Set to Null when CapturaAntibody should be diluted into samples (in FastELISA).
Default Calculation: If Method is set to DirectSandwichELISA or IndirectSandwichELISA, the option is automatically set to Model[Sample, StockSolution, "1x Carbonate-Bicarbonate Buffer pH10"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
CaptureAntibodyStorageCondition
Default Calculation: If Method is set to DirectSandwichELISA, IndirectSandwichELISA, or FastELISA, Automatically set to Refrigerator.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
ReferenceAntigen
The sample containing the antigen that is used in DirectCompetitiveELISA or IndirectCompetitiveELISA. The ReferenceAntigen competes with TargetAntigen in the samples for the binding of the PrimaryAntibody. Reference Antigen is also referred to as Inhibitor Antigen.
Default Calculation: Automatically set to a sample containing known amount of TargetAntigen when Method is set to DirectCompetitiveELISA or IndirectCompetitiveELISA.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
ReferenceAntigenDilutionFactor
The dilution ratio of ReferenceAntigen. The ReferenceAntigenSample is always diluted in ReferenceAntigenDiluent. Either ReferenceAntigenDilutionFactor or ReferenceAntigenVolume should be provided but not both.
Default Calculation: If Method is DirectCompetitiveELISA or IndirectCompetitiveELISA, automatically set to 0.001 (1:1,000).
ReferenceAntigenVolume
The volume of undiluted ReferenceAntigen added into the corresponding well of the assay plate. ReferenceAntigenVolume is used as an alternative to ReferenceAntigenDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
ReferenceAntigenDiluent
Default Calculation: Method is DirectCompetitiveELISA and IndirectCompetitiveELISA, the option is automatically set to Model[Sample, StockSolution, "1x Carbonate-Bicarbonate Buffer pH10"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
ReferenceAntigenStorageCondition
Default Calculation: Automatically set to Refrigerator if Method is DirectCompetitiveELISA and IndirectCompetitiveELISA.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
PrimaryAntibody
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
PrimaryAntibodyDilutionFactor
The dilution ratio of PrimaryAntibody. For DirectELISA, IndirectELISA, DirectSandwichELISA, and IndirectSandwichELISA, the antibody is diluted with PrimaryAntibodyDiluent. For DirectCompetitiveELISA, IndirectCompetitiveELISA, and FastELISA, the antibody is diluted with the corresponding sample. Either PrimaryAntibodyDilutionFactor or PrimaryAntibodyVolume should be provided but not both.
Default Calculation: If used for DirectELISA, IndirectELISA, DirectSandwichELISA, IndirectSandwichELISA, automatically set to 0.001 (1:1,000). If used for DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, automatically set to 0.01 (1:100).
PrimaryAntibodyVolume
The volume of undiluted PrimaryAntibody added into the corresponding well of the assay plate. PrimaryAntibodyVolume is used as an alternative to PrimaryAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
PrimaryAntibodyDiluent
The buffer used to dilute the PrimaryAntibody.Set to Null when PrimaryAntibody should be diluted into samples (in DirectCompetitiveELISA, IndirectCompetitiveELISA, and FastELISA).
Default Calculation: If Method is set to DirectELISA, IndirectELISA, DirectSandwichELISA, IndirectSandwichELISA, the options will be automatically set to Model[Sample,"ELISA Blocker Blocking Buffer"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
PrimaryAntibodyStorageCondition
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal.
SecondaryAntibody
Default Calculation: If Method is IndirectELISA, IndirectSandwichELISA, or IndirectCompetitiveELISA, the option is automatically set to a stocked secondary antibody for the primary antibody.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
SecondaryAntibodyDilutionFactor
The dilution ratio of SecondaryAntibody. SecondaryAntibody is always diluted in the SecondryAntibodyDiluent. Either SecondaryAntibodyDilutionFactor or SecondaryAntibodyVolume should be provided but not both.
Default Calculation: For IndirectELISA, IndirectSandwichELISA, IndirectCompetitiveELISA, automatically set to 0.001 (1:1,000).
SecondaryAntibodyVolume
The volume of SecondaryAntibody added into the corresponding well of the assay plate. SecondaryAntibodyVolume is used as an alternative to SecondaryAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
SecondaryAntibodyDiluent
Default Calculation: If Method is set to IndirectELISA, IndirectSandwichELISA, IndirectCompetitiveELISA, the option will automatically set to Model[Sample,"ELISA Blocker Blocking Buffer"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
SecondaryAntibodyStorageCondition
Default Calculation: If method is IndirectELISA, IndirectSandwichELISA, or IndirectCompetitiveELISA, the option is automatically set to Refrigerator.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
Sample Antibody Complex Incubation
SampleAntibodyComplexIncubation
Indicates if the pre-mixed samples and antibodies should be incubated before loaded into the assay plate. The plate used for sample-antibody mixing will be discarded after the experiment.
Default Calculation: Automatically set to True if Method is set to DirectCompetitiveELISA, IndirectCompetitiveELISA, and FastELISA.
SampleAntibodyComplexIncubationTime
The duration of sample-antibody complex incubation (If needed). In DirectCompetitiveELISA and IndirectCompetitiveELISA, PrimaryAntibody is incubated with the sample. In FastELISA, PrimaryAntibody and CaptureAntibody is incubated with the sample. If Null, the prepared sample-antibody complex will be kept at 4 degree Celsius till ready to use.
Default Calculation: If the Method is set to DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, the option is automatically set to 2 Hours.
SampleAntibodyComplexIncubationTemperature
The temperature at which sample mixed with Antibodies are incubated. In DirectCompetitiveELISA and IndirectCompetitiveELISA, PrimaryAntibody is incubated with the sample. In FastELISA, PrimaryAntibody and CaptureAntibody are incubated with the sample. If Null, the prepared sample-antibody complex will be used directly.
Default Calculation: If the Method is set to DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, the option is automatically set to Ambient.
Pattern Description: Ambient or greater than or equal to 4 degrees Celsius and less than or equal to 50 degrees Celsius or Null.
Coating
Coating
Indicates if Coating is required. Coating is a procedure to non-specifically adsorb protein molecules to the surface of wells of the assay plate.
SampleCoatingVolume
The amount of Sample that is aliquoted into the assay plate, in order for the Sample to be adsorbed to the surface of the well.
Default Calculation: If Method is DirectELISA or IndirectELISA, CoatingVolume is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
CoatingAntibodyCoatingVolume
The amount of diluted CoatingAntibody that is aliquoted into the assay plate, in order for the CoatingAntibody to be adsorbed to the surface of the well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 300 microliters or Null.
ReferenceAntigenCoatingVolume
The amount of diluted ReferenceAntigen that is aliquoted into the assay plate, in order for the ReferenceAntigen to be adsorbed to the surface of the well.
Default Calculation: If Method is DirectCompetitiveELISA or IndirectCompetitiveELISA, CoatingVolume is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
CaptureAntibodyCoatingVolume
The amount of diluted CaptureAntibody that is aliquoted into the assay plate, in order for the CaptureAntibody to be adsorbed to the surface of the well.
Default Calculation: If Method is DirectSandwichELISA or IndirectSandwichELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
CoatingTemperature
The temperature at which the Coating Solution is kept in the assay plate, in order for the coating molecules to be adsorbed to the surface of the well.
Default Calculation: If Coating is set to True, the option is automatically set to 4 degree Celsius.
Pattern Description: Ambient or greater than or equal to 4 degrees Celsius and less than or equal to 50 degrees Celsius or Null.
CoatingTime
The duration when the Coating Solution is kept in the assay plate, in order for the coating molecules to be adsorbed to the surface of the well.
CoatingWashVolume
The volume of WashBuffer added to rinse off unbound molecule. When Coating is False but Blocking is True, the pre-coated plate must be washed at least once to ensure no liquid is left in the plate.
Default Calculation: If Coating is set to True, or Coating is False but Blocking is True, the option is automatically set to 250 Microliters.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 300 microliters or Null.
CoatingNumberOfWashes
The number of washes performed after coating. When Coating is False but Blocking is True, the pre-coated plate must be washed at least once to ensure no liquid is left in the plate.
Default Calculation: If Coating is set to True,or Coating is False but Blocking is True, the option is automatically set to 3.
Blocking
Blocking
Indicates if a protein solution should be incubated with the assay plate to prevent non-specific binding of molecules to the assay plate.
BlockingBuffer
The protein-containing solution used to prevent non-specific binding of antigen or antibody to the surface of the assay plate.
Default Calculation: If Blocking is True, automatically set to Model[Sample,"ELISA Blocker Blocking Buffer"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlockingVolume
The amount of BlockingBuffer that is aliquoted into the appropriate wells of the assay plate, in order to prevent non-specific binding of molecules to the assay plate.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 300 microliters or Null.
BlockingTime
The duration when the BlockingBuffer is kept with the assay plate, in order to prevent non-specific binding of molecules to the assay plate.
BlockingTemperature
The duration of time when the BlockingBuffer is kept with the assay plate, in order to prevent non-specific binding of molecules to the assay plate.
Pattern Description: Greater than or equal to 25 degrees Celsius and less than or equal to 50 degrees Celsius or Null.
BlockingMixRate
The speed at which the plate is shaken (orbitally, at a radius of 2 mm) during Blocking incubation. Mixing is not recommended when incubation volume is high, in which case the option should be set to Null.
Pattern Description: Greater than or equal to 40 revolutions per minute and less than or equal to 1000 revolutions per minute or Null.
BlockingWashVolume
The volume of WashBuffer added after Blocking, in order to rinse off the unbound molecules from the surface of the wells. If Coating and Blocking are both False, at least one blocking wash is required to ensure no liquid is remaining in the assay plate.
Default Calculation: If Blocking is True, or Coating and Blocking are both False, the option is automatically set to 250 Microliters.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 300 microliters or Null.
BlockingNumberOfWashes
The number of washes performed after Blocking, in order to rinse off the unbound molecules from the surface of the wells.
Default Calculation: If Blocking is True, or Coating and Blocking are both False, the option is automatically set to 3.
Immunosorbent Step
SampleAntibodyComplexImmunosorbentVolume
The volume of the sample-antibody complex to be loaded on each well of the ELISAPlate. In DirectCompetitiveELISA and IndirectCompetitiveELISA, this step enables the free primary antibody to bind to the ReferenceAntigen coated on the plate. In FastELISA, this step enables the PrimaryAntibody-TargetAntigen-CaptureAntibody complex to bind to the CoatingAntibody on the plate.
Default Calculation: If the Method is set to DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
SampleAntibodyComplexImmunosorbentTime
Default Calculation: If the Method is set to DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, the option is automatically set to 2 Hour.
SampleAntibodyComplexImmunosorbentTemperature
Default Calculation: If the Method is set to DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, the option is automatically set to 25 Celsius.
Pattern Description: Ambient or greater than or equal to 4 degrees Celsius and less than or equal to 50 degrees Celsius or Null.
SampleAntibodyComplexImmunosorbentMixRate
The speed at which the plate is shaken (orbitally, at a radius of 2 mm) during SampleAntibody mixture incubation in the assay plate. Mixing is not recommended when incubation volume is higher than 200 Microliters, in which case the options should be set to Null.
Pattern Description: Greater than or equal to 40 revolutions per minute and less than or equal to 1000 revolutions per minute or Null.
SampleAntibodyComplexImmunosorbentWashVolume
The volume of WashBuffer added to rinse off the unbound primary antibody after sample-antibody complex incubation.
Default Calculation: If the Method is set to DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, the option is automatically set to 250 Microliter.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 300 microliters or Null.
SampleAntibodyComplexImmunosorbentNumberOfWashes
Default Calculation: If the Method is set to DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, the option is automatically set to 4.
SampleImmunosorbentVolume
The volume of the Sample to be loaded on the ELISAPlate for the target antigen to bind to the capture antibody in DirectSandwichELISA and IndirectSandwichELISA.
Default Calculation: If the Method is set to DirectSandwichELISA and IndirectSandwichELISA, the option is automatically set to 100 Microliters.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
SampleImmunosorbentTime
Default Calculation: If the Method is set to DirectSandwichELISA and IndirectSandwichELISA, the option is automatically set to 2 Hour.
SampleImmunosorbentTemperature
Default Calculation: If the Method is set to DirectSandwichELISA and IndirectSandwichELISA, the option is automatically set to 25 Celsius.
Pattern Description: Greater than or equal to 25 degrees Celsius and less than or equal to 50 degrees Celsius or Null.
SampleImmunosorbentMixRate
The speed at which the plate is shaken (orbitally, at a radius of 2 mm) during Sample incubation. Mixing is not recommended when incubation volume is higher than 200 Microliters, in which case the options should be set to Null.
Pattern Description: Greater than or equal to 40 revolutions per minute and less than or equal to 1000 revolutions per minute or Null.
SampleImmunosorbentWashVolume
Default Calculation: If the Method is set to DirectSandwichELISA and IndirectSandwichELISA, the option is automatically set to 250 Microliters.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 300 microliters or Null.
SampleImmunosorbentNumberOfWashes
Default Calculation: If the Method is set to DirectSandwichELISA and IndirectSandwichELISA, the option is automatically set to 4.
PrimaryAntibodyImmunosorbentVolume
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
PrimaryAntibodyImmunosorbentTime
Default Calculation: If the Method is set to DirectELISA, IndirectELISA, DirectSandwichELISA, or IndirectSandwichELISA, the option is automatically set to 2 Hour.
PrimaryAntibodyImmunosorbentTemperature
Default Calculation: If the Method is set to DirectELISA, IndirectELISA, DirectSandwichELISA, or IndirectSandwichELISA, the option is automatically set to 25 Celsius.
Pattern Description: Greater than or equal to 25 degrees Celsius and less than or equal to 50 degrees Celsius or Null.
PrimaryAntibodyImmunosorbentMixRate
The speed at which the plate is shaken (orbitally, at a radius of 2 mm) during PrimaryAntibody incubation. Mixing is not recommended when incubation volume is higher than 200 Microliters, in which case the options should be set to Null.
Pattern Description: Greater than or equal to 40 revolutions per minute and less than or equal to 1000 revolutions per minute or Null.
PrimaryAntibodyImmunosorbentWashVolume
The volume of WashBuffer added to rinse off the unbound primary antibody after PrimaryAntibody incubation.
Default Calculation: If the Method is set to DirectELISA, IndirectELISA, DirectSandwichELISA, or IndirectSandwichELISA, the option is automatically set to 250 Microliters.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 300 microliters or Null.
PrimaryAntibodyImmunosorbentNumberOfWashes
Default Calculation: If the Method is set to DirectELISA, IndirectELISA, DirectSandwichELISA, or IndirectSandwichELISA, the option is automatically set to 4.
SecondaryAntibodyImmunosorbentVolume
Default Calculation: If the Method is set to IndirectELISA, IndirectSandwichELISA, and IndirectCompetitiveELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
SecondaryAntibodyImmunosorbentTime
Default Calculation: If the Method is set to IndirectELISA, IndirectSandwichELISA, and IndirectCompetitiveELISA, the option is automatically set to 1 Hour.
SecondaryAntibodyImmunosorbentTemperature
Default Calculation: If the Method is set to IndirectELISA, IndirectSandwichELISA, and IndirectCompetitiveELISA, the option is automatically set to 25 Celsius.
Pattern Description: Greater than or equal to 25 degrees Celsius and less than or equal to 50 degrees Celsius or Null.
SecondaryAntibodyImmunosorbentMixRate
The speed at which the plate is shaken (orbitally, at a radius of 2 mm) during Secondary Antibody incubation. Mixing is not recommended when incubation volume is higher than 200 Microliters, in which case the options should be set to Null.
Pattern Description: Greater than or equal to 40 revolutions per minute and less than or equal to 1000 revolutions per minute or Null.
SecondaryAntibodyImmunosorbentWashVolume
The volume of WashBuffer added to rinse off the unbound Secondary antibody after SecondaryAntibody incubation.
Default Calculation: If the Method is set to IndirectELISA, IndirectSandwichELISA, and IndirectCompetitiveELISA, the option is automatically set to 250 Microliters.
Pattern Description: Greater than or equal to 25 microliters and less than or equal to 300 microliters or Null.
SecondaryAntibodyImmunosorbentNumberOfWashes
Default Calculation: If the Method is set to IndirectELISA, IndirectSandwichELISA, and IndirectCompetitiveELISA, the option is automatically set to 4.
Detection
SubstrateSolution
Default Calculation: If enzyme is Horseredish Peroxidase, the option will be automatically set to Model[Sample,"ELISA TMB Stabilized Chromogen"]. If enzyme is Alkaline Phosphatase, then the option is automatically set to Model[Sample,"AP Substrate PNPP Solution"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample.
StopSolution
Default Calculation: If enzyme is Horseradish Peroxidase, then the option is automatically set to Model[Sample,"ELISA HRP-TMB Stop Solution"]. If enzyme is Alkaline Phosphatase, then the option is automatically set to Model[Sample,"AP Substrate Stop Solution"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
SubstrateSolutionVolume
Default Calculation: If SubstrateSolution is populated, then the option is automatically set to 100ul.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters.
StopSolutionVolume
Default Calculation: If StopSolution is selected, the StopSolutionVolume is automatically set to 100ul.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
SubstrateIncubationTime
SubstrateIncubationTemperature
The temperature of the Substrate incubation, in order for the detection reaction to take place where the antibody-conjugated enzyme (such as Horseradish Peroxidase or Alkaline Phosphatase) catalyzes the colorimetric reaction.
Pattern Description: Greater than or equal to 25 degrees Celsius and less than or equal to 50 degrees Celsius.
SubstrateIncubationMixRate
The speed at which the plate is shaken (orbitally, at a radius of 2 mm) during Substrate incubation. Mixing is not recommended when incubation volume is higher than 200 Microliters, in which case the options should be set to Null.
Pattern Description: Greater than or equal to 40 revolutions per minute and less than or equal to 1000 revolutions per minute or Null.
AbsorbanceWavelength
Pattern Description: {405 nanometers, 450 nanometers, 492 nanometers, 620 nanometers} or list of one or more {405 nanometers, 450 nanometers, 492 nanometers, 620 nanometers} entries.
Programmatic Pattern: (405*Nanometer | 450*Nanometer | 492*Nanometer | 620*Nanometer) | {(405*Nanometer | 450*Nanometer | 492*Nanometer | 620*Nanometer)..}
PrereadBeforeStop
Indicate if colorimetric reactions between the enzyme and its substrate will be checked by the plate reader before the stopping reagent was added to terminate the reaction.
Default Calculation: Is set automatically to True if either or both of PrereadTimepoints and PrereadAbsorbanceWavelength are specified. Otherwise is set to False.
PrereadTimepoints
The list of time points when absorbance intensitied at each PreparedAbsorbanceWavelength during the Preread process (before the colorimetric reaction was terminated).
Default Calculation: If PrereadBeforeStop is True, this option is set to one single value: the half of the SubstrateIncubationTime. Otherwise is set to Null.
Pattern Description: Greater than or equal to 0 minutes and less than or equal to 72 hours or list of one or more greater than or equal to 0 minutes and less than or equal to 72 hours entries or Null.
Programmatic Pattern: ((RangeP[0*Minute, $MaxExperimentTime] | {RangeP[0*Minute, $MaxExperimentTime]..}) | Automatic) | Null
PrereadAbsorbanceWavelength
The wavelength used to detect the absorbance of light during the Preread process (before the colorimetric reaction was terminated).
Default Calculation: If PrereadBeforeStop is True, this option is set to {450 Nanometer}. Otherwise is set to Null.
Pattern Description: {405 nanometers, 450 nanometers, 492 nanometers, 620 nanometers} or list of one or more {405 nanometers, 450 nanometers, 492 nanometers, 620 nanometers} entries or Null.
Programmatic Pattern: (((405*Nanometer | 450*Nanometer | 492*Nanometer | 620*Nanometer) | {(405*Nanometer | 450*Nanometer | 492*Nanometer | 620*Nanometer)..}) | Automatic) | Null
SignalCorrection
The absorbance reading that is used to eliminate the interference of background absorbance (such as from ELISAPlate material and dust). If True, a reading at 620 nm is read at the same time of the AbsorbanceWavelength. The correction is done by subtracting the reading at 620nm from that at the AbsorbanceWavelength.
Standard
Standard
A sample containing known amount of TargetAntigen molecule. Standard is used for the quantification of Standard analyte.
Default Calculation: If TargetAntigen is specified, standard will be automatically set to the DefaultSampleModel of the target antigen.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardTargetAntigen
The Analyte molecule(e.g., peptide, protein, and hormone) detected and quantified in the Standard samples by Antibodies in the ELISA experiment. This option is used to automatically set sample Antibodies and the corresponding experiment conditions of Standards and Blanks.
StandardSerialDilutionCurve
The collection of serial dilutions that will be performed on sample. StandardLoadingVolume of each dilution will be transferred to the ELISAPlate. For Serial Dilution Volumes, the Transfer Volume is taken out of the sample and added to a second well with the Diluent Volume of the Diluent. It is mixed, then the Transfer Volume is taken out of that well to be added to a third well. This is repeated to make Number Of Dilutions diluted samples. For example, if a 100 ug/ ml sample with a Transfer Volume of 20 Microliters, a Diluent Volume of 60 Microliters and a Number of Dilutions of 3 is used, it will create a DilutionCurve of 25 ug/ ml, 6.25 ug/ ml, and 1.5625 ug/ ml with each dilution having a volume of 60 Microliters. For Serial Dilution Factors, the sample will be diluted by the dilution factor at each transfer step. IMPORTANT: because the dilution curve does not intrinsically include the original sample, if the original standard solution is to be included in the dilution curve, the first diluting factor should be set to 1 or Diluent Volume should be 0 Microliters. During experiment, an 5% extra volume than specified is going to be prepared in order to offset pipetting errors. Therefore, the total volume for each well specified in this option should be equal or greater than the volume needed for the experiment. Because the dilution of each assay (well) is prepared independently, the NumberOfReplications should not be considered when determining the Standard Volume. The plate used for sample dilutions will be discarded after the experiment. Note: if spike or antibodies are to be mixed with samples, their volumes are counted towards diluent volume.
Figure 3.3: Use the SerialDilutionCuve option to create a collection of serial dilutions that will be performed on standards.
Default Calculation: Automatically set to Null if the StandardDilutionCurve is specified. In all other cases it is automatically set to a Standard Volume of 100 Microliters and a Constant Dilution Factor of 0.5. The Number Of Dilutions is automatically set to 6.
Programmatic Pattern: (({GreaterEqualP[0*Microliter], {RangeP[0, 1], GreaterP[0, 1]} | {RangeP[0, 1]..}} | {RangeP[0*Microliter, 1000*Microliter], RangeP[40*Microliter, 1000*Microliter], GreaterP[0, 1]}) | Automatic) | Null
StandardDilutionCurve
The collection of dilutions that will be performed on each sample. For Fixed Dilution Volume Dilution Curves, the Standard Amount is the volume of the sample that will be mixed the Diluent Volume of the Diluent to create a desired concentration. The Standard Volume is the TOTAL volume of the sample that will created after being diluted by the Dilution Factor. For example, a 1 ug/ ml sample with a dilution factor of 0.7 will be diluted to a concentration 0.7 ug/ ml. IMPORTANT: because the dilution curve does not intrinsically include the original sample, if the original standard solution is to be included in the dilution curve, the first diluting factor should be set to 1 or Diluent Volume must be 0 Microliter. During experiment, an 5% extra volume than specified is going to be prepared in order to offset pipetting errors. Therefore, the total volume for each well specified in this option should be equal or greater than the volume needed for the experiment. Because the dilution of each assay (well) is prepared independently, the NumberOfReplications should not be considered when determining the Standard Volume. The plate used for sample dilutions will be discarded after the experiment. Note: if spike or antibodies are to be mixed with samples, their volumes are counted towards diluent volume.
Figure 3.4: Use the DilutionCuve option to create a collection of dilutions that will be performed on standards.
Default Calculation: Automatically set to Null if the SerialDilutionCurve is specified. If SerialDilutionCurve is set to Null, then automatically set Standard Assay Volume to 100ul, dilution factor to 0.5, 0,25, 0.125, 0.0625, 0.03125, 0.015625.
Programmatic Pattern: (({{RangeP[0*Microliter, 1000*Microliter], RangeP[0*Microliter, 1000*Microliter]}..} | {{RangeP[40*Microliter, 1500*Microliter], RangeP[0, 1]}..}) | Automatic) | Null
StandardDiluent
Default Calculation: If StandardDilutionCurve and StandardSerialDilutionCurve are not both Null, in the case when Method is set to DirectELISA and IndirectELISA, the option is automatically set to Model[Sample, StockSolution, "1x Carbonate-Bicarbonate Buffer pH10"]; in all other cases, the option is automatically set to Model[Sample,"ELISA Blocker Blocking Buffer"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardStorageCondition
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
StandardCoatingAntibody
Default Calculation: If Method is FastELISA and Standard is not Null, automatically set to an antibody against a tag which is conjugated to a the StandardCaptureAntibody.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardCoatingAntibodyDilutionFactor
The dilution ratio of StandardCoatingAntibody. StandardCoatingAntibody is diluted with CoatingAntibodyDiluent. Either StandardCoatingAntibodyDilutionFactor or StandardCoatingAntibodyVolume should be provided but not both.
Default Calculation: If Method is FastELISA and Standard is not Null, automatically set to 0.001 (1:1,000)
StandardCoatingAntibodyVolume
The volume of undiluted StandardCoatingAntibody added into the corresponding well of the assay plate. StandardCoatingAntibody is diluted with CoatingAntibodyDiluent. StandardCoatingAntibodyVolume is used as an alternative to StandardCoatingAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardCaptureAntibody
The sample containing the antibody that is used to pull down the antigen from sample solution to the plate in DirectSandwichELISA, IndirectSandwichELISA, and FastELISA.
Default Calculation: Automatically set to Null is Standard is Null. If Method is FastELISA, automatically set to an antibody containing an affinity tag and against the TargetAntigen. If Method is DirectSandwichELISA or IndirectSandwichELISA, automatically set to an un-tagged antibody against TargetAntigen.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardCaptureAntibodyDilutionFactor
The dilution ratio of StandardCaptureAntibody. For DirectSandwichELISA and IndirectSandwichELISA, StandardCaptureAntibody is diluted with CaptureAntibodyDiluent. For FastELISA, StandardCaptureAntibody is diluted in the corresponding standard. Either StandardCaptureAntibodyDilutionFactor or StandardCaptureAntibodyVolume should be provided but not both.
Default Calculation: When Standard is not Null, if Method is DirectSandwichELISA or IndirectSandwichELISA, automatically set to 0.001 (1:1,000). If Method is FastELISA, automatically set to 0.01 (1:100).
StandardCaptureAntibodyVolume
The volume of undiluted CaptureAntibody added into the corresponding well of the assay plate. StandardCaptureAntibodyVolume is used as an alternative to StandardCaptureAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardReferenceAntigen
The sample containing the antigen that is used in DirectCompetitiveELISA or IndirectCompetitiveELISA. The StandardReferenceAntigen competes with sample antigen for the binding of the StandardPrimaryAntibody. Reference antigen is sometimes also referred to as inhibitor antigen.
Default Calculation: Automatically set to a sample containing known amount of TargetAntigen when Method is set to DirectCompetitiveELISA or IndirectCompetitiveELISA and Standard is not Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardReferenceAntigenDilutionFactor
The dilution ratio of StandardReferenceAntigen. For DirectCompetitiveELISA and IndirectCompetitiveELISA, the StandardReferenceAntigenStandard is diluted in ReferenceAntigenDiluent. Either StandardReferenceAntigenDilutionFactor or StandardReferenceAntigenVolume should be provided but not both.
Default Calculation: Automatically set to 0.001 (1:1,000) if Method is DirectCompetitiveELISA or IndirectCompetitiveELISA.
StandardReferenceAntigenVolume
The volume of ReferenceAntigen added into the corresponding well of the assay plate. StandardReferenceAntigenVolume is used as an alternative to StandardReferenceAntigenDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardPrimaryAntibody
Default Calculation: If Standard is not Null, the option will be automatically set to an antibody against the TargetAntigen.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardPrimaryAntibodyDilutionFactor
The dilution ratio of StandardPrimaryAntibody. For DirectELISA, IndirectELISA, DirectSandwichELISA, and IndirectSandwichELISA, the antibody is diluted with PrimaryAntibodyDiluent. For DirectCompetitiveELISA, IndirectCompetitiveELISA, and FastELISA, the antibody is diluted in the corresponding sample. Either StandardPrimaryAntibodyDilutionFactor or StandardPrimaryAntibodyVolume should be provided but not both.
Default Calculation: When Standard is not Null, if used for DirectELISA, IndirectELISA, DirectSandwichELISA, IndirectSandwichELISA, automatically set to 0.001 (1:1,000). If used for DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, automatically set to 0.01 (1:100).
StandardPrimaryAntibodyVolume
The volume of PrimaryAntibody added into the corresponding well of the assay plate. StandardPrimaryAntibodyVolume is used as an alternative to StandardPrimaryAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardSecondaryAntibody
Default Calculation: If Method is IndirectELISA, IndirectSandwichELISA, or IndirectCompetitiveELISA, the option is automatically set to a stocked secondary antibody for the primary antibody.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardSecondaryAntibodyDilutionFactor
The dilution ratio of StandardSecondaryAntibody. StandardSecondaryAntibody is always diluted in the SecondaryAntibodyDiluent. Either StandardSecondaryAntibodyDilutionFactor or StandardSecondaryAntibodyVolume should be provided but not both.
Default Calculation: If Standard is not Null, for IndirectELISA, IndirectSandwichELISA, IndirectCompetitiveELISA, automatically set to 0.001 (1:1,000).
StandardSecondaryAntibodyVolume
The volume of SecondaryAntibody added into the corresponding well of the assay plate. StandardSecondaryAntibodyVolume is used as an alternative to StandardSecondaryAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardCoatingVolume
The amount of Standard that is aliquoted into the ELISAPlate, in order for the Standard to be adsorbed to the surface of the well.
Default Calculation: If Method is DirectELISA or IndirectELISA, StandardCoatingVolume is automatically set to 200 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardReferenceAntigenCoatingVolume
The amount of StandardReferenceAntigen that is aliquoted into the assay plate, in order for the StandardReferenceAntigen to be adsorbed to the surface of the well.
Default Calculation: If Method is DirectCompetitiveELISA or IndirectCompetitiveELISA, CoatingVolume is automatically set to 200 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardCoatingAntibodyCoatingVolume
The amount of diluted StandardCoatingAntibody that is aliquoted into the ELISAPlate, in order for the StandardCoatingAntibody to be adsorbed to the surface of the well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardCaptureAntibodyCoatingVolume
The amount of diluted StandardCaptureAntibody that is aliquoted into the ELISAPlate, in order for the StandardCaptureAntibody to be adsorbed to the surface of the well.
Default Calculation: If Method is DirectSandwichELISA or IndirectSandwichELISA, the option is automatically set to 200 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardBlockingVolume
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 300 microliters or Null.
StandardAntibodyComplexImmunosorbentVolume
The volume of the standard-antibody complex to be loaded on the ELISAPlate. In DirectCompetitiveELISA and IndirectCompetitiveELISA, this step enables the free StandardPrimaryAntibody to bind to the StandardReferenceAntigen coated on the plate. In FastELISA, this step enables the PrimaryAntibody-TargetAntigen-CaptureAntibody complex to bind to the CoatingAntibody on the plate.
Default Calculation: If the Method is set to DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardImmunosorbentVolume
The volume of the Standard to be loaded on the ELISAPlate for the target antigen to bind to the capture antibody in DirectSandwichELISA and IndirectSandwichELISA.
Default Calculation: If the Method is set to DirectSandwichELISA and IndirectSandwichELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardPrimaryAntibodyImmunosorbentVolume
Default Calculation: If the Method is set to DirectELISA, IndirectELISA, DirectSandwichELISA, or IndirectSandwichELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardSecondaryAntibodyImmunosorbentVolume
Default Calculation: If the Method is set to IndirectELISA, IndirectSandwichELISA, and IndirectCompetitiveELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardSubstrateSolution
Default Calculation: If enzyme is Horseredish Peroxidase, the option will be automatically set to Model[Sample,"ELISA TMB Stabilized Chromogen"]. If enzyme is Alkaline Phosphatase, then the option is automatically set to Model[Sample,"AP Substrate PNPP Solution"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardStopSolution
Default Calculation: If enzyme is Horseradish Peroxidase, then the option is automatically set to Model[Sample,"ELISA HRP-TMB Stop Solution"]. If enzyme is Alkaline Phosphatase, then the option is automatically set to Model[Sample,"AP Substrate Stop Solution"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
StandardSubstrateSolutionVolume
Default Calculation: If StandardSubstrateSolution is populated and Standard is not Null, then the option is automatically set to 100ul.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
StandardStopSolutionVolume
Default Calculation: If Model[Sample,"ELISA TMB Stabilized Chromogen"] or Model[Sample,"ELISA HRP-TMB Stop Solution"] is selected for StandardSubstrateSolution, the StopSolutionVolume is automatically set to 100ul.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
Blank
Blank
Default Calculation: If In DirectELISA and IndirectELISA, the option will be automatically set to Model[Sample, StockSolution, "1x Carbonate-Bicarbonate Buffer pH10"]. Otherwise, the option will be automatically set to Model[Sample, "ELISA Blocker Blocking Buffer"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlankTargetAntigen
The Analyte molecule(e.g., peptide, protein, and hormone) detected and quantified in the blanks by Antibodies in the ELISA experiment. This option is used to automatically set Antibodies and the corresponding experiment conditions.
BlankStorageCondition
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
BlankCoatingAntibody
Default Calculation: If Method is FastELISA and Blank is not Null, automatically set to an antibody against a tag which is conjugated to a the StandardCaptureAntibody.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlankCoatingAntibodyDilutionFactor
The dilution ratio of BlankCoatingAntibody. BlankCoatingAntibody is diluted with CoatingAntibodyDiluent. Either BlankCoatingAntibodyDilutionFactor or BlankCoatingAntibodyVolume should be provided but not both.
Default Calculation: If Method is FastELISA and Blank is not Null, automatically set to 0.001 (1:1,000)
BlankCoatingAntibodyVolume
The volume of BlankCoatingAntibody added into the corresponding well of the assay plate. BlankCoatingAntibody is diluted with CoatingAntibodyDiluent. BlankCoatingAntibodyVolume is used as an alternative to BlankCoatingAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankCaptureAntibody
The sample containing the antibody that is used to pull down the antigen from sample solution to the plate in DirectSandwichELISA, IndirectSandwichELISA, and FastELISA.
Default Calculation: Automatically set to Null is Blank is Null. If Method is FastELISA, automatically set to an antibody containing an affinity tag and against the TargetAntigen. If Method is DirectSandwichELISA or IndirectSandwichELISA, automatically set to an un-tagged antibody against TargetAntigen.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlankCaptureAntibodyDilutionFactor
The dilution ratio of BlankCaptureAntibody. For DirectSandwichELISA and IndirectSandwichELISA, BlankCaptureAntibody is diluted with CaptureAntibodyDiluent. For FastELISA, BlankCaptureAntibody is diluted in the corresponding sample. Either BlankCaptureAntibodyDilutionFactor or BlankCaptureAntibodyVolume should be provided but not both.
Default Calculation: If Method is DirectSandwichELISA or IndirectSandwichELISA, automatically set to 0.001 (1:1,000). If Method is FastELISA, automatically set to 0.01 (1:100).
BlankCaptureAntibodyVolume
The volume of CaptureAntibody added into the corresponding well of the assay plate. BlankCaptureAntibodyVolume is used as an alternative to BlankCaptureAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankReferenceAntigen
The Sample containing the antigen that is used in DirectCompetitiveELISA or IndirectCompetitiveELISA. The BlankReferenceAntigen competes with sample antigen for the binding of the BlankPrimaryAntibody. Reference antigen is sometimes also referred to as inhibitor antigen.
Default Calculation: Automatically set to a sample containing known amount of TargetAntigen when Method is set to DirectCompetitiveELISA or IndirectCompetitiveELISA and Blank is not Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlankReferenceAntigenDilutionFactor
The dilution ratio of BlankReferenceAntigen. For DirectCompetitiveELISA and IndirectCompetitiveELISA, the BlankReferenceAntigenBlank is diluted in ReferenceAntigenDiluent. Either BlankReferenceAntigenDilutionFactor or BlankReferenceAntigenVolume should be provided but not both.
Default Calculation: Automatically set to 0.001 (1:1,000) if Method is DirectCompetitiveELISA or IndirectCompetitiveELISA.
BlankReferenceAntigenVolume
The volume of ReferenceAntigen added into the corresponding well of the assay plate. BlankReferenceAntigenVolume is used as an alternative to BlankReferenceAntigenDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankPrimaryAntibody
Default Calculation: The option will be automatically set to an antibody against the BlankTargetAntigen if Blank is not Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlankPrimaryAntibodyDilutionFactor
The dilution ratio of BlankPrimaryAntibody. For DirectELISA, IndirectELISA, DirectSandwichELISA, and IndirectSandwichELISA, the antibody is diluted with PrimaryAntibodyDiluent. For DirectCompetitiveELISA, IndirectCompetitiveELISA, and FastELISA, the antibody is diluted in the corresponding sample. Either BlankPrimaryAntibodyDilutionFactor or BlankPrimaryAntibodyVolume should be provided but not both.
Default Calculation: When Blank is not Null, if used for DirectELISA, IndirectELISA, DirectSandwichELISA, IndirectSandwichELISA, automatically set to 0.001 (1:1,000). If used for DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, automatically set to 0.01 (1:100).
BlankPrimaryAntibodyVolume
The volume of PrimaryAntibody added into the corresponding well of the assay plate. BlankPrimaryAntibodyVolume is used as an alternative to BlankPrimaryAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankSecondaryAntibody
Default Calculation: If Method is IndirectELISA, IndirectSandwichELISA, or IndirectCompetitiveELISA and Blank is not Null, the option is automatically set to an stocked secondary antibody for the primary antibody.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlankSecondaryAntibodyDilutionFactor
The dilution ratio of BlankSecondaryAntibody. BlankSecondaryAntibody is always diluted in the SecondaryAntibodyDiluent. Either BlankSecondaryAntibodyDilutionFactor or BlankSecondaryAntibodyVolume should be provided but not both.
Default Calculation: If Blank is not Null, for IndirectELISA, IndirectSandwichELISA, IndirectCompetitiveELISA, automatically set to 0.001 (1:1,000).
BlankSecondaryAntibodyVolume
The volume of BlankSecondaryAntibody added into the corresponding well of the assay plate. BlankSecondaryAntibodyVolume is used as an alternative to BlankSecondaryAntibodyDilutionFactor. During antibody preparation, a master mix will be made for antibody dilution, and the diluted Antibodies will be aliquoted into each well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankCoatingVolume
Default Calculation: If Blank is not Null, and Method is DirectELISA or IndirectELISA, CoatingVolume is automatically set to 200 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankReferenceAntigenCoatingVolume
The amount of diluted BlankReferenceAntigen that is aliquoted into the assay plate, in order for the BlankReferenceAntigen to be adsorbed to the surface of the well.
Default Calculation: If Method is DirectCompetitiveELISA or IndirectCompetitiveELISA, CoatingVolume is automatically set to 200 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankCoatingAntibodyCoatingVolume
The amount of diluted BlankCoatingAntibody that is aliquoted into the ELISAPlate, in order for the BlankCoatingAntibody to be adsorbed to the surface of the well.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankCaptureAntibodyCoatingVolume
The amount of diluted BlankCaptureAntibody that is aliquoted into the ELISAPlate, in order for the BlankCaptureAntibody to be adsorbed to the surface of the well.
Default Calculation: If Method is DirectSandwichELISA or IndirectSandwichELISA, the option is automatically set to 200 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankBlockingVolume
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 300 microliters or Null.
BlankAntibodyComplexImmunosorbentVolume
The volume of the BlankAntibodyComplex to be loaded on the ELISAPlate. In DirectCompetitiveELISA and IndirectCompetitiveELISA, this step enables the free primary antibody to bind to the ReferenceAntigen coated on the plate. In FastELISA, this step enables the PrimaryAntibody-TargetAntigen-CaptureAntibody complex to bind to the CoatingAntibody on the plate.
Default Calculation: If the Method is set to DirectCompetitiveELISA, IndirectCompetitiveELISA, or FastELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankImmunosorbentVolume
The volume of the Blank to be loaded on the ELISAPlate for the target antigen to bind to the capture antibody in DirectSandwichELISA and IndirectSandwichELISA.
Default Calculation: If the Method is set to DirectSandwichELISA and IndirectSandwichELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankPrimaryAntibodyImmunosorbentVolume
Default Calculation: If the Method is set to DirectELISA, IndirectELISA, DirectSandwichELISA, or IndirectSandwichELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankSecondaryAntibodyImmunosorbentVolume
Default Calculation: If the Method is set to IndirectELISA, IndirectSandwichELISA, and IndirectCompetitiveELISA, the option is automatically set to 100 Microliter.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankSubstrateSolution
Default Calculation: If enzyme is Horseredish Peroxidase, the option will be automatically set to Model[Sample,"ELISA TMB Stabilized Chromogen"]. If enzyme is Alkaline Phosphatase, then the option is automatically set to Model[Sample,"AP Substrate PNPP Solution"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlankStopSolution
Default Calculation: If enzyme is Horseradish Peroxidase, then the option is automatically set to Model[Sample,"ELISA HRP-TMB Stop Solution"]. If enzyme is Alkaline Phosphatase, then the option is automatically set to Model[Sample,"AP Substrate Stop Solution"].
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BlankSubstrateSolutionVolume
Default Calculation: If SubstrateSolution is populated, then the option is automatically set to 100ul.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
BlankStopSolutionVolume
Default Calculation: If Model[Sample,"ELISA TMB Stabilized Chromogen"] or Model[Sample,"ELISA HRP-TMB Stop Solution"] is selected, the StopSolutionVolume is automatically set to 100ul.
Pattern Description: Greater than or equal to 0 microliters and less than or equal to 200 microliters or Null.
Assay Plate
ELISAPlate
The assay plate each sample, standard, and blank will be loaded into and the immunosorbent assay will take place. This plate can be pre-coated and blocked in advance.
Default Calculation: The option will be automatically set to an optical, flat-bottom 96 well plate made with polystyrene. If Samples are pre-coated to the a plate, the option will be automatically set to this plate.
Pattern Description: An object of type or subtype Model[Container, Plate] or Object[Container, Plate] or a prepared sample.
Programmatic Pattern: (ObjectP[{Model[Container, Plate], Object[Container, Plate]}] | _String) | Automatic
SecondaryELISAPlate
Default Calculation: If the total number of Sample (including spiked samples and dilutions), standards, and blanks, times NumberOfReplications exceeds 96, then the option will be automatically set to an optical, flat-bottom 96 well plate made with polystyrene. If Samples are pre-coated to two plates, the option will be automatically set to the second plate.
Pattern Description: An object of type or subtype Model[Container, Plate] or Object[Container, Plate] or a prepared sample or Null.
Programmatic Pattern: ((ObjectP[{Model[Container, Plate], Object[Container, Plate]}] | _String) | Automatic) | Null
ELISAPlateAssignment
Specifies the placement of samples and their corresponding spikes, dilutions, and antibodies in the first ELISAPlate.
Default Calculation: Samples are automatically assigned according to the input sequence (UnknownSamples,Standards,Blanks), to the ELISAPlate from the A1 position, consecutively, in a column-wise fashion. Each sample and their dilution curve will be placed next to each other, then each replicates are placed next to each other. When samples are pre-coated onto the plate, they are arranged from the A1 position, consequtively, in a column-wise fashion,regardless of the input sequence. If the first plate cannot accommodate all the samples, the samples will automatically be assigned to the SecondaryELISAPlate. Due to possible plate-to-plate variations, when two plates are needed, users are recommended to rearrange the samples so that the samples, standards, and blanks that are detected using the same conditions (i.e. same experiment) should be placed in the same plate. If impossible, additional standards and blanks should be included so that each sample has a corresponding set of standards and blanks on the same plate.
Pattern Description: List of one or more {Type, Sample, Spike, SpikeDilutionFactor, SampleDilutionFactors, CoatingAntibody, CoatingAntibodyDilutionFactor, CaptureAntibody, CaptureAntibodyDilutionFactor, ReferenceAntigen, ReferenceAntigenDilutionFactor, PrimaryAntibody, PrimaryAntibodyDilutionFactor, SecondaryAntibody, SecondaryAntibodyDilutionFactor, CoatingVolume, BlockingVolume, SampleAntibodyComplexImmunosorbentVolume, SampleImmunosorbentVolume, PrimaryAntibodyImmunosorbentVolume, SecondaryAntibodyImmunosorbentVolume, SubstrateSolution, StopSolution, SubstrateSolutionVolume, StopSolutionVolume} entries.
Programmatic Pattern: {{Unknown | Spike | Standard | Blank, ObjectP[{Model[Sample], Object[Sample]}] | _String, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Null, RangeP[0, 1] | Null, {RangeP[0, 1]..}, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Null, RangeP[0, 1] | Null, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Null, RangeP[0, 1] | Null, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Null, RangeP[0, 1] | Null, ObjectP[{Model[Sample], Object[Sample]}] | _String, RangeP[0, 1] | Null, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Null, RangeP[0, 1] | Null, RangeP[0*Microliter, 200*Microliter] | Null, RangeP[0*Microliter, 300*Microliter] | Null, RangeP[0*Microliter, 200*Microliter] | Null, RangeP[0*Microliter, 200*Microliter] | Null, RangeP[0*Microliter, 200*Microliter] | Null, RangeP[0*Microliter, 200*Microliter] | Null, ObjectP[{Model[Sample], Object[Sample]}] | _String, ObjectP[{Model[Sample], Object[Sample]}] | _String, RangeP[0*Microliter, 200*Microliter] | Null, RangeP[0*Microliter, 200*Microliter] | Null}..} | Automatic
SecondaryELISAPlateAssignment
Specifies the placement of samples and their corresponding spikes, dilutions, and antibodies in the second ELISAPlate.
Default Calculation: Samples are automatically assigned according to the input sequence (UnknownSamples,Standards,Blanks), to the ELISAPlate from the A1 position, consecutively, in a column-wise fashion. Each sample and their dilution curve will be placed next to each other, then each replicates are placed next to each other. When samples are pre-coated onto the plate, they are arranged from the A1 position, consequtively, in a column-wise fashion,regardless of the input sequence. If the first plate cannot accommodate all the samples, the samples will automatically be assigned to the SecondaryELISAPlate. Due to possible plate-to-plate variations, when two plates are needed, users are recommended to rearrange the samples so that the samples, standards, and blanks that are detected using the same conditions (i.e. same experiment) should be placed in the same plate. If impossible, additional standards and blanks should be included so that each sample has a corresponding set of standards and blanks on the same plate.
Pattern Description: List of one or more {Type, Sample, Spike, SpikeDilutionFactor, SampleDilutionFactors, CoatingAntibody, CoatingAntibodyDilutionFactor, CaptureAntibody, CaptureAntibodyDilutionFactor, ReferenceAntigen, ReferenceAntigenDilutionFactor, PrimaryAntibody, PrimaryAntibodyDilutionFactor, SecondaryAntibody, SecondaryAntibodyDilutionFactor, CoatingVolume, BlockingVolume, SampleAntibodyComplexImmunosorbentVolume, SampleImmunosorbentVolume, PrimaryAntibodyImmunosorbentVolume, SecondaryAntibodyImmunosorbentVolume, SubstrateSolution, StopSolution, SubstrateSolutionVolume, StopSolutionVolume} entries or Null.
Programmatic Pattern: ({{Unknown | Spike | Standard | Blank, ObjectP[{Model[Sample], Object[Sample]}] | _String, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Null, RangeP[0, 1] | Null, {RangeP[0, 1]..}, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Null, RangeP[0, 1] | Null, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Null, RangeP[0, 1] | Null, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Null, RangeP[0, 1] | Null, ObjectP[{Model[Sample], Object[Sample]}] | _String, RangeP[0, 1] | Null, (ObjectP[{Model[Sample], Object[Sample]}] | _String) | Null, RangeP[0, 1] | Null, RangeP[0*Microliter, 200*Microliter] | Null, RangeP[0*Microliter, 300*Microliter] | Null, RangeP[0*Microliter, 200*Microliter] | Null, RangeP[0*Microliter, 200*Microliter] | Null, RangeP[0*Microliter, 200*Microliter] | Null, RangeP[0*Microliter, 200*Microliter] | Null, ObjectP[{Model[Sample], Object[Sample]}] | _String, ObjectP[{Model[Sample], Object[Sample]}] | _String, RangeP[0*Microliter, 200*Microliter] | Null, RangeP[0*Microliter, 200*Microliter] | Null}..} | Automatic) | Null
Model Input
PreparedModelContainer
Indicates the container in which a Model[Sample] specified as input to the experiment function will be prepared.
Default Calculation: If PreparedModelAmount is set to All and when the input model has a product associated with both Amount and DefaultContainerModel populated, automatically set to the DefaultContainerModel value in the product. Otherwise set to Model[Container, Vessel, "2mL Tube"].
PreparedModelAmount
Indicates the amount of a Model[Sample] specified as input to the experiment function that will be prepared in the PreparedModelContainer. When set to All and the input model sample is not preparable, the entire amount of the input model sample that comes from one of the Products is prepared. The selected product must have both Amount and DefaultContainerModel populated, and it must not be a KitProduct. When set to All and the input model is preparable such as water, 1 Milliliter of the input model sample is prepared.
Programmatic Pattern: ((RangeP[1*Microliter, 20*Liter] | RangeP[1*Milligram, 20*Kilogram] | GreaterP[0*Unit, 1*Unit] | GreaterP[0., 1.] | All) | Automatic) | Null
Post Experiment
SamplesInStorageCondition
The non-default conditions under which the SamplesIn of this experiment should be stored after the protocol is completed. If left unset, SamplesIn will be stored according to their current StorageCondition.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
Sample Prep Options
Sample Preparation
PreparatoryUnitOperations
Specifies a sequence of transferring, aliquoting, consolidating, or mixing of new or existing samples before the main experiment. These prepared samples can be used in the main experiment by referencing their defined name. For more information, please reference the documentation for ExperimentSamplePreparation.
Pattern Description: List of one or more unit Operation ManualSamplePreparation or RoboticSamplePreparation or unit Operation must match SamplePreparationP entries or Null.
Programmatic Pattern: {((ManualSamplePreparationMethodP | RoboticSamplePreparationMethodP) | SamplePreparationP)..} | Null
Preparatory Incubation
Incubate
Indicates if the SamplesIn should be incubated at a fixed temperature prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Incubation options are set. Otherwise, resolves to False.
IncubationTemperature
Temperature at which the SamplesIn should be incubated for the duration of the IncubationTime prior to starting the experiment.
Pattern Description: Ambient or greater than or equal to -20 degrees Celsius and less than or equal to 500 degrees Celsius or Null.
Programmatic Pattern: ((Ambient | RangeP[$MinIncubationTemperature, $MaxIncubationTemperature]) | Automatic) | Null
IncubationTime
Duration for which SamplesIn should be incubated at the IncubationTemperature, prior to starting the experiment.
Mix
Default Calculation: Automatically resolves to True if any Mix related options are set. Otherwise, resolves to False.
MixType
Default Calculation: Automatically resolves based on the container of the sample and the Mix option.
Pattern Description: Roll, Vortex, Sonicate, Pipette, Invert, Stir, Shake, Homogenize, Swirl, Disrupt, or Nutate or Null.
MixUntilDissolved
Indicates if the mix should be continued up to the MaxIncubationTime or MaxNumberOfMixes (chosen according to the mix Type), in an attempt dissolve any solute. Any mixing/incubation will occur prior to starting the experiment.
Default Calculation: Automatically resolves to True if MaxIncubationTime or MaxNumberOfMixes is set.
MaxIncubationTime
Maximum duration of time for which the samples will be mixed while incubated in an attempt to dissolve any solute, if the MixUntilDissolved option is chosen. This occurs prior to starting the experiment.
Default Calculation: Automatically resolves based on MixType, MixUntilDissolved, and the container of the given sample.
IncubationInstrument
Default Calculation: Automatically resolves based on the options Mix, Temperature, MixType and container of the sample.
Pattern Description: An object of type or subtype Model[Instrument, Roller], Model[Instrument, OverheadStirrer], Model[Instrument, Vortex], Model[Instrument, Shaker], Model[Instrument, BottleRoller], Model[Instrument, Roller], Model[Instrument, Sonicator], Model[Instrument, HeatBlock], Model[Instrument, Homogenizer], Model[Instrument, Disruptor], Model[Instrument, Nutator], Model[Instrument, Thermocycler], Model[Instrument, EnvironmentalChamber], Model[Instrument, Pipette], Object[Instrument, Roller], Object[Instrument, OverheadStirrer], Object[Instrument, Vortex], Object[Instrument, Shaker], Object[Instrument, BottleRoller], Object[Instrument, Roller], Object[Instrument, Sonicator], Object[Instrument, HeatBlock], Object[Instrument, Homogenizer], Object[Instrument, Disruptor], Object[Instrument, Nutator], Object[Instrument, Thermocycler], Object[Instrument, EnvironmentalChamber], or Object[Instrument, Pipette] or Null.
AnnealingTime
Minimum duration for which the SamplesIn should remain in the incubator allowing the system to settle to room temperature after the IncubationTime has passed but prior to starting the experiment.
IncubateAliquotContainer
The desired type of container that should be used to prepare and house the incubation samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
IncubateAliquotDestinationWell
The desired position in the corresponding IncubateAliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
IncubateAliquot
The amount of each sample that should be transferred from the SamplesIn into the IncubateAliquotContainer when performing an aliquot before incubation.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
Preparatory Centrifugation
Centrifuge
Indicates if the SamplesIn should be centrifuged prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Centrifuge options are set. Otherwise, resolves to False.
CentrifugeInstrument
Pattern Description: An object of type or subtype Model[Instrument, Centrifuge] or Object[Instrument, Centrifuge] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, Centrifuge], Object[Instrument, Centrifuge]}] | Automatic) | Null
CentrifugeIntensity
The rotational speed or the force that will be applied to the samples by centrifugation prior to starting the experiment.
Pattern Description: Greater than 0 revolutions per minute or greater than 0 standard accelerations due to gravity on the surface of the earth or Null.
Programmatic Pattern: ((GreaterP[0*RPM] | GreaterP[0*GravitationalAcceleration]) | Automatic) | Null
CentrifugeTime
CentrifugeTemperature
The temperature at which the centrifuge chamber should be held while the samples are being centrifuged prior to starting the experiment.
Pattern Description: Ambient or greater than or equal to -10 degrees Celsius and less than or equal to 40 degrees Celsius or Null.
CentrifugeAliquotContainer
The desired type of container that should be used to prepare and house the centrifuge samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
CentrifugeAliquotDestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
CentrifugeAliquot
The amount of each sample that should be transferred from the SamplesIn into the CentrifugeAliquotContainer when performing an aliquot before centrifugation.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
Preparatory Filtering
Filtration
Indicates if the SamplesIn should be filter prior to starting the experiment or any aliquoting. Sample Preparation occurs in the order of Incubation, Centrifugation, Filtration, and then Aliquoting (if specified).
Default Calculation: Resolves to True if any of the corresponding Filter options are set. Otherwise, resolves to False.
FiltrationType
Default Calculation: Will automatically resolve to a filtration type appropriate for the volume of sample being filtered.
FilterInstrument
Default Calculation: Will automatically resolved to an instrument appropriate for the filtration type.
Pattern Description: An object of type or subtype Model[Instrument, FilterBlock], Object[Instrument, FilterBlock], Model[Instrument, PeristalticPump], Object[Instrument, PeristalticPump], Model[Instrument, VacuumPump], Object[Instrument, VacuumPump], Model[Instrument, Centrifuge], Object[Instrument, Centrifuge], Model[Instrument, SyringePump], or Object[Instrument, SyringePump] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, FilterBlock], Object[Instrument, FilterBlock], Model[Instrument, PeristalticPump], Object[Instrument, PeristalticPump], Model[Instrument, VacuumPump], Object[Instrument, VacuumPump], Model[Instrument, Centrifuge], Object[Instrument, Centrifuge], Model[Instrument, SyringePump], Object[Instrument, SyringePump]}] | Automatic) | Null
Filter
The filter that should be used to remove impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Will automatically resolve to a filter appropriate for the filtration type and instrument.
Pattern Description: An object of type or subtype Model[Container, Plate, Filter], Model[Container, Vessel, Filter], or Model[Item, Filter] or Null.
Programmatic Pattern: (ObjectP[{Model[Container, Plate, Filter], Model[Container, Vessel, Filter], Model[Item, Filter]}] | Automatic) | Null
FilterMaterial
The membrane material of the filter that should be used to remove impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Resolves to an appropriate filter material for the given sample is Filtration is set to True.
Pattern Description: Cellulose, Cotton, Polyethylene, Polypropylene, PTFE, Nylon, PES, PLUS, PVDF, GlassFiber, GHP, UHMWPE, EPDM, DuraporePVDF, GxF, ZebaDesaltingResin, NickelResin, AgaroseResin, CobaltResin, Silica, HLB, or AnoporeAlumina or Null.
PrefilterMaterial
The material from which the prefilter filtration membrane should be made of to remove impurities from the SamplesIn prior to starting the experiment.
Pattern Description: Cellulose, Cotton, Polyethylene, Polypropylene, PTFE, Nylon, PES, PLUS, PVDF, GlassFiber, GHP, UHMWPE, EPDM, DuraporePVDF, GxF, ZebaDesaltingResin, NickelResin, AgaroseResin, CobaltResin, Silica, HLB, or AnoporeAlumina or Null.
FilterPoreSize
The pore size of the filter that should be used when removing impurities from the SamplesIn prior to starting the experiment.
Default Calculation: Resolves to an appropriate filter pore size for the given sample is Filtration is set to True.
Pattern Description: 0.008 micrometers, 0.02 micrometers, 0.1 micrometers, 0.2 micrometers, 0.22 micrometers, 0.45 micrometers, 1. micrometer, 1.1 micrometers, 2.5 micrometers, 6. micrometers, 20. micrometers, 30. micrometers, or 100. micrometers or Null.
PrefilterPoreSize
The pore size of the filter; all particles larger than this should be removed during the filtration.
Pattern Description: 0.008 micrometers, 0.02 micrometers, 0.1 micrometers, 0.2 micrometers, 0.22 micrometers, 0.45 micrometers, 1. micrometer, 1.1 micrometers, 2.5 micrometers, 6. micrometers, 20. micrometers, 30. micrometers, or 100. micrometers or Null.
FilterSyringe
Default Calculation: Resolves to an syringe appropriate to the volume of sample being filtered, if Filtration is set to True.
Pattern Description: An object of type or subtype Model[Container, Syringe] or Object[Container, Syringe] or a prepared sample or Null.
Programmatic Pattern: ((ObjectP[{Model[Container, Syringe], Object[Container, Syringe]}] | _String) | Automatic) | Null
FilterHousing
The filter housing that should be used to hold the filter membrane when filtration is performed using a standalone filter membrane.
Default Calculation: Resolve to an housing capable of holding the size of the membrane being used, if filter with Membrane FilterType is being used and Filtration is set to True.
Pattern Description: An object of type or subtype Model[Instrument, FilterHousing], Object[Instrument, FilterHousing], Model[Instrument, FilterBlock], or Object[Instrument, FilterBlock] or Null.
Programmatic Pattern: (ObjectP[{Model[Instrument, FilterHousing], Object[Instrument, FilterHousing], Model[Instrument, FilterBlock], Object[Instrument, FilterBlock]}] | Automatic) | Null
FilterIntensity
Default Calculation: Will automatically resolve to 2000 GravitationalAcceleration if FiltrationType is Centrifuge and Filtration is True.
Pattern Description: Greater than 0 revolutions per minute or greater than 0 standard accelerations due to gravity on the surface of the earth or Null.
Programmatic Pattern: ((GreaterP[0*RPM] | GreaterP[0*GravitationalAcceleration]) | Automatic) | Null
FilterTime
Default Calculation: Will automatically resolve to 5 Minute if FiltrationType is Centrifuge and Filtration is True.
FilterTemperature
The temperature at which the centrifuge chamber will be held while the samples are being centrifuged during filtration.
Default Calculation: Will automatically resolve to 22 Celsius if FiltrationType is Centrifuge and Filtration is True.
FilterContainerOut
The desired container filtered samples should be produced in or transferred into by the end of filtration, with indices indicating grouping of samples in the same plates, if desired.
Default Calculation: Automatically set as the PreferredContainer for the Volume of the sample. For plates, attempts to fill all wells of a single plate with the same model before using another one.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or {Index, Container} or Null.
Programmatic Pattern: (((ObjectP[{Model[Container], Object[Container]}] | _String) | {GreaterEqualP[1, 1] | Automatic, (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
FilterAliquotDestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
FilterAliquotContainer
The desired type of container that should be used to prepare and house the filter samples which should be used in lieu of the SamplesIn for the experiment.
Programmatic Pattern: ((ObjectP[Model[Container]] | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | Automatic}) | Automatic) | Null
FilterAliquot
The amount of each sample that should be transferred from the SamplesIn into the FilterAliquotContainer when performing an aliquot before filtration.
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container.
Pattern Description: All or greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
FilterSterile
Default Calculation: Resolve to False if Filtration is indicated. If sterile filtration is desired, this option must manually be set to True.
Aliquoting
Aliquot
Indicates if aliquots should be taken from the SamplesIn and transferred into new AliquotSamples used in lieu of the SamplesIn for the experiment. Note that if NumberOfReplicates is specified this indicates that the input samples will also be aliquoted that number of times. Note that Aliquoting (if specified) occurs after any Sample Preparation (if specified).
AliquotAmount
Default Calculation: Automatically set as the smaller between the current sample volume and the maximum volume of the destination container if a liquid, or the current Mass or Count if a solid or counted item, respectively.
Programmatic Pattern: ((RangeP[1*Microliter, 20*Liter] | RangeP[1*Milligram, 20*Kilogram] | GreaterP[0*Unit, 1*Unit] | GreaterP[0., 1.] | All) | Automatic) | Null
TargetConcentration
The desired final concentration of analyte in the AliquotSamples after dilution of aliquots of SamplesIn with the ConcentratedBuffer and BufferDiluent which should be used in lieu of the SamplesIn for the experiment.
TargetConcentrationAnalyte
Default Calculation: Automatically set to the first value in the Analytes field of the input sample, or, if not populated, to the first analyte in the Composition field of the input sample, or if none exist, the first identity model of any kind in the Composition field.
Pattern Description: An object of type or subtype Model[Molecule], Model[Molecule, cDNA], Model[Molecule, Oligomer], Model[Molecule, Transcript], Model[Molecule, Protein], Model[Molecule, Protein, Antibody], Model[Molecule, Carbohydrate], Model[Molecule, Polymer], Model[Resin], Model[Resin, SolidPhaseSupport], Model[Lysate], Model[ProprietaryFormulation], Model[Virus], Model[Cell], Model[Cell, Mammalian], Model[Cell, Bacteria], Model[Cell, Yeast], Model[Tissue], Model[Material], or Model[Species] or Null.
AssayVolume
Default Calculation: Automatically determined based on Volume and TargetConcentration option values.
Pattern Description: Greater than or equal to 1 microliter and less than or equal to 20 liters or Null.
ConcentratedBuffer
The concentrated buffer which should be diluted by the BufferDilutionFactor in the final solution (i.e., the combination of the sample, ConcentratedBuffer, and BufferDiluent). The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
BufferDilutionFactor
The dilution factor by which the concentrated buffer should be diluted in the final solution (i.e., the combination of the sample, ConcentratedBuffer, and BufferDiluent). The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: If ConcentratedBuffer is specified, automatically set to the ConcentratedBufferDilutionFactor of that sample; otherwise, set to Null.
BufferDiluent
The buffer used to dilute the aliquot sample such that ConcentratedBuffer is diluted by BufferDilutionFactor in the final solution. The ConcentratedBuffer and BufferDiluent will be combined and then mixed with the sample, where the combined volume of these buffers is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: Automatically resolves to Model[Sample, "Milli-Q water"] if ConcentratedBuffer is specified; otherwise, resolves to Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
AssayBuffer
The buffer that should be added to any aliquots requiring dilution, where the volume of this buffer added is the difference between the AliquotAmount and the total AssayVolume.
Default Calculation: Automatically resolves to Model[Sample, "Milli-Q water"] if ConcentratedBuffer is not specified; otherwise, resolves to Null.
Pattern Description: An object of type or subtype Model[Sample] or Object[Sample] or a prepared sample or Null.
AliquotSampleStorageCondition
The non-default conditions under which any aliquot samples generated by this experiment should be stored after the protocol is completed.
Pattern Description: {AmbientStorage, EnclosedAmbientStorage, Refrigerator, Freezer, DeepFreezer, CryogenicStorage, YeastIncubation, YeastShakingIncubation, BacterialIncubation, BacterialShakingIncubation, MammalianIncubation, ViralIncubation, CrystalIncubation, AcceleratedTesting, IntermediateTesting, LongTermTesting, UVVisLightTesting} or Disposal or Null.
DestinationWell
The desired position in the corresponding AliquotContainer in which the aliquot samples will be placed.
Default Calculation: Automatically resolves to A1 in containers with only one position. For plates, fills wells in the order provided by the function AllWells.
Pattern Description: Any well from A1 to H12 or list of one or more any well from A1 to H12 or any well from A1 to H12 entries or Null.
Programmatic Pattern: ((WellPositionP | {((Automatic | Null) | WellPositionP)..}) | Automatic) | Null
AliquotContainer
The desired type of container that should be used to prepare and house the aliquot samples, with indices indicating grouping of samples in the same plates, if desired. This option will resolve to be the length of the SamplesIn * NumberOfReplicates.
Default Calculation: Automatically set as the PreferredContainer for the AssayVolume of the sample. For plates, attempts to fill all wells of a single plate with the same model before aliquoting into the next.
Pattern Description: An object of type or subtype Model[Container] or Object[Container] or a prepared sample or Automatic or Null or {Index, Container} or list of one or more an object of type or subtype Model[Container] or Object[Container] or a prepared sample or Automatic or Null entries or list of one or more Automatic or Null or {Index, Container} entries.
Programmatic Pattern: (((ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null) | {GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null)} | {((ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null))..} | {({GreaterEqualP[1, 1] | (Automatic | Null), (ObjectP[{Model[Container], Object[Container]}] | _String) | (Automatic | Null)} | (Automatic | Null))..}) | Automatic) | Null
AliquotPreparation
Default Calculation: Automatic resolution will occur based on manipulation volumes and container types.
ConsolidateAliquots
Protocol Options
Organizational Information
Template
A template protocol whose methodology should be reproduced in running this experiment. Option values will be inherited from the template protocol, but can be individually overridden by directly specifying values for those options to this Experiment function.
Pattern Description: An object of type or subtype Object[Protocol] or an object of type or subtype of Object[Protocol] with UnresolvedOptions, ResolvedOptions specified or Null.
Programmatic Pattern: (ObjectP[Object[Protocol]] | FieldReferenceP[Object[Protocol], {UnresolvedOptions, ResolvedOptions}]) | Null
Name
A object name which should be used to refer to the output object in lieu of an automatically generated ID number.
Post Experiment
MeasureWeight
Indicates if any solid samples that are modified in the course of the experiment should have their weights measured and updated after running the experiment. Please note that public samples are weighed regardless of the value of this option.
MeasureVolume
Indicates if any liquid samples that are modified in the course of the experiment should have their volumes measured and updated after running the experiment. Please note that public samples are volume measured regardless of the value of this option.
ImageSample
Example Calls
ELISA Method
DirectELISA method detects the immobilized antigen by an antibody directly conjugated with an enzyme:
IndirectELISA method detects the immobilized antigen by an unlabled primary antibody and a secondary antibody conjugated to an enzyme:
DirectSandwichELISA method first applies the antigen to a plate with pre-coated capture antibody before detecting in a DirectELISA configuration:
IndirectSandwichELISA method first applies the antigen to a plate with pre-coated capture antibody before detecting in a IndirectELISA configuration:
DirectCompetitiveELISA method detects antigen by allowing the antigen to competes with a reference for binding to labeled antibodies in a DirectELISA configuration:
IndirectCompetitiveELISA method detects antigen by allowing the antigen to competes with a reference for binding to unlabeled antibodies in a IndirectELISA configuration:
FastELISA method detects antigen by using a coated antibody against a tag which is conjugated to another antigen-targeting antibody:
Sample Spiking
The input samples can be mixed with the desired spike samples with known concentration(s) of analyte(s), for evaluation of sample matrices in a new ELISA assay:
Sample Dilution
A serial dilution curve can be specified for an input sample by providing the volumes of the diluted samples, a constant dilution factor, and a number of dilutions. This should be used if you want the sample to be diluted by the same factor at each step of the serial dilution. The volume of the sample will be the final volume of the sample after the dilution is performed. The example call below will create a series of samples with dilution factors of (0.1,0.01) and a volume of 100 Microliters:
A serial dilution curve can be specified for an input sample by providing the volumes of the diluted samples and each dilution factor.This should be used if you want the sample to be diluted by different factors at each step of the serial dilution. The volume of the sample will be the final volume of the sample after the dilution is performed. The example call below will create a series of samples with dilution factors of (0.5, 0.05, 0.005) and a volume of 100 Microliters:
A serial dilution curve can be specified for an input sample by providing the volume of sample to be transferred, the volume of diluent and the number of dilutions. This should be used if you want give the volumes the sample will be diluted by with instead of dilution factors. The example call below will create a series of samples with dilution factors of (0.9, 0.81) and a volume of 10 Microliters:
A dilution curve can be specified for an input sample by providing the volumes of the diluted samples and the dilution factor:
Standard and Blank
Standard samples can be added to ELISA experiment to provide information about the accuracy of the assay or be used for the generation of the standard curves:
Blank samples can be added to ELISA experiment to measure the background signal from the reagents and the instrument:
Antibodies and Reference Antigen
SecondaryAntibody-- to specify secondary antibody that binds to the primary antibody-antigen complex
CaptureAntibody (for DierectSandwichELISA and IndirectSandwichELISA)-- to specify the antibody coated on the ELISA Plate that pulls down the target antigen from samples.
ReferenceAntigen-- to specify the sample containing the target antigen coated on the ELISA Plate that competes with the antigen in the sample for the binding of the primary antibody.
Warnings and Errors
Messages (59)
Within the same option, an Object[Sample] and its Model[Sample] cannot coexist:
Reagents used for Standard and Blank must be also exist for samples:
Within the same option, an Object[Sample] and its Model[Sample] cannot coexist:
AntibodyVolumeIncompatibility (1)
CaptureAntibodyPipettingVolumeTooLow (1)
CoatingAntibodyPipettingVolumeTooLow (1)
DilutionCurveOptionsTotalVolumeTooLarge (1)
DilutionCurvesPipettingVolumeTooLow (1)
EitherOrConflictingOptions (2)
The options (if index-matched, matched to the same parent), should have one and only one specified. Use PrimaryAntibodyDilutionFactor and PrimaryAntibodyVolume as example:
The dilution factor and dilution volume options (if index-matched, matched to the same parent), should have one and only one specified. Use PrimaryAntibodyDilutionFactor and PrimaryAntibodyVolume as example:
ElisaImproperContainers (1)
ElisaIncompatibleContainer (1)
ELISAInvalidPrereadOptions (1)
ELISAInvalidPrereadTimepoints (1)
IndexedSameNullConflictingOptions (2)
The index-matched options with the same parent (e.g., SubstrateSolution and SubstrateSolutionVolume) should be all Null or all Unull at the same time:
The index-matched options with the same parent (e.g., SpikeDilutionFactor and SpikeStorageCondition) should be all Null or all Unull at the same time:
MasterConflictingNullOptions (6)
When the master switch option (Method) is set to DirectSandwichELISA, the related single options (e.g., PrimaryAntibodyDiluent) cannot be set to Null:
When the master switch option SampleAntibodyComplexIncubation is set to True, the related single options cannot be set to Null:
When the master switch option Coating is set to True, the related single options cannot be set to Null:
When the master switch option (Method) is set to DirectSandwichELISA, the related index matching options (e.g., SampleImmunosorbentVolume) cannot be set to Null:
When the master switch option Blocking is set to True, the related index matching options cannot be set to Null:
When the master switch option Coating is set to True and the option Method is set to a certain value (e.g., FastELISA), the related index matching options (dilution and coating options) must also be set to Null:
MasterConflictingUnullOptions (11)
When the master switch option (e.g., Standard) is set to Null, all corresponding single options to Null:
When the master switch option (e.g., Blank) is set to Null, all corresponding single options to Null:
When the master switch option Method is set to a certain value (e.g., DirectCompetitiveELISA), the unused antibody single options must be set to Null:
When the master switch option SampleAntibodyComplexIncubation is set to False, the related single options must also be set to Null:
When the master switch option Coating is set to False, the related single options must also be set to Null:
When the master switch option Coating is set to False and the option Method is set to a certain value (e.g., DirectSandwichELISA), the related diluent option must also be set to Null:
When the master switch option (e.g., Standard) is set to Null, all corresponding index matched options to Null:
When the master switch option (e.g., Blank) is set to Null, all corresponding inex matched options to Null:
When the master switch option Method is set to a certain value (e.g., IndirectELISA), the unused antibody index matching options must be set to Null:
When the master switch option Blocking is set to False, the related index matching options must also be set to Null:
When the master switch option Coating is set to False and the option Method is set to a certain value (e.g., IndirectELISA), the related index matching options (dilution and coating options) must also be set to Null:
Measure620WithCorrection (1)
MethodConflictingSampleAntibodyIncubationSwitch (1)
NotMoreThanOneConflictingOptions (1)
ObjectDoesNotExist (6)
Throw a message if we have a sample that does not exist (name form):
Throw a message if we have a container that does not exist (name form):
Throw a message if we have a sample that does not exist (ID form):
Throw a message if we have a container that does not exist (ID form):
Do NOT throw a message if we have a simulated sample but a simulation is specified that indicates that it is simulated:
Do NOT throw a message if we have a simulated container but a simulation is specified that indicates that it is simulated:
PlateAssignmentDoesNotMatchOptions (2)
PrecoatedSamplesCannotBeAliquoted (1)
PrimaryAntibodyPipettingVolumeTooLow (1)
ReferenceAntigenPipettingVolumeTooLow (1)
SampleVolumeShouldNotBeLargerThan50ml (2)
SecondaryAntibodyPipettingVolumeTooLow (1)
SpecifyDiluentAndDilutionCurveTogether (1)
SpecifySpikeWithFixedDilutionCurve (1)
SpikePipettingVolumeTooLow (1)
UnresolvableIndexMatchedOptions (2)
With the provided information, required options should be automatically resolved. PrimaryAntibody is resolved from the presented analyte in the sample:
With the provided information, required options should be automatically resolved. SubstrateSolution is resolved from the enzyme conjugated to the secondary antibody:
Possible Issues
Evaporation during prolonged incubation
All incubations after ELISAPlate coating are performed in a plate incubator without plate seals. Prolonged incubation at high temperature (such as 37 °C for 3 hours) may result in evaporation of samples. To reduce such effect, avoid long incubation of samples with very small volumes (such as under 50 Microliter) at a high temperature.
Uneven incubation time
The 4-channel pipette loads liquid into 4 wells at a time. When incubation time is short but number of wells is large, the time-difference between the loading of first and last wells may affect data. For this, we recommend incubation times to be longer than 30 minutes.
Sub-optimal Dilution
Dilutions of samples, standards, and antibodies should be empirically optimized for the best performance of the experiment. For example, if antibody is over dilute, the signal may be too weak to be detected.
Sub-optimal Diluent
Diluent for samples, standards and antibodies should be empirically optimized for the best performance of the experiment. Form example, biocarbonate buffer at PH 10 may result in better coating than phosphate-buffered saline at PH 6.7.
Detection reagent
Last modified on Sat 16 Aug 2025 15:35:05