The IRF (Interferon Regulatory Factor) ELISA Kit is designed for the quantitative measurement of Interferon Regulatory Factor (IRF) proteins in biological samples such as cell lysates, tissue homogenates, and possibly serum or plasma. IRFs are a family of transcription factors that play critical roles in regulating immune responses, including the activation of genes involved in antiviral defense, immune cell development, and inflammation. The most studied IRFs include IRF3, IRF7, and IRF9, each involved in different aspects of immune regulation.
Content of the Kit
- Pre-coated 96-Well Microplate:
- Wells pre-coated with an antibody specific to IRF, allowing capture of the IRF protein from the samples.
- Standards:
- Solutions containing known concentrations of recombinant or purified IRF proteins, used to create a standard curve for quantifying IRF levels in samples.
- Sample Diluent:
- A buffer solution used to dilute biological samples to appropriate concentrations for accurate measurement.
- Detection Antibody:
- An antibody specific to a different epitope of IRF, often conjugated to an enzyme such as HRP for detection.
- Enzyme Conjugate:
- In some kits, the detection antibody may be biotinylated, requiring a streptavidin-HRP conjugate for signal detection.
- Substrate Solution (TMB):
- Tetramethylbenzidine, which reacts with HRP to produce a colorimetric signal proportional to the amount of IRF.
Assay Procedure Summary
- Sample and Standard Addition: Add standards and samples to the microplate wells where IRF proteins are captured by the immobilized antibody.
- Wash: Remove unbound substances by washing the wells.
- Detection Antibody Addition: Add a detection antibody specific to IRF, which binds to the captured protein.
- Wash: Wash away excess detection antibody.
- Enzyme Conjugate Addition: If the detection antibody is biotinylated, add a streptavidin-HRP conjugate to bind to the biotinylated antibody.
- Wash: Remove unbound enzyme conjugate through washing.
Performance Characteristics
- Sensitivity: Capable of detecting low levels of IRF proteins, typically in the picogram per milliliter (pg/mL) range.
- Precision: Low intra-assay and inter-assay variability, with coefficients of variation (CV) generally below 10%.
- Linearity: Demonstrates linearity across the standard curve range, ensuring accurate quantification.
- Recovery: High recovery rates, usually above 85%, indicating reliable performance across various sample types.
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