A TCR (T-cell receptor) ELISA (Enzyme-Linked Immunosorbent Assay) Kit is used for the quantitative measurement of T-cell receptor levels in various biological samples, such as serum, plasma, cell culture supernatants, and tissue homogenates. TCRs are critical components of the adaptive immune system, recognizing antigens presented by MHC (Major Histocompatibility Complex) molecules and playing a pivotal role in immune responses.
Content of the Kit
- Pre-coated 96-Well Microplate:
- Wells are coated with a monoclonal antibody specific to the TCR of interest (e.g., anti-TCR α/β or γ/δ chains).
- Standards:
- Calibrated solutions containing known concentrations of TCR, used to generate a standard curve for quantification.
- Sample Diluent:
- A buffer used to dilute samples to optimal concentrations for accurate measurement.
- Detection Antibody:
- Biotinylated or enzyme-conjugated antibody specific to a different epitope on the TCR, ensuring specific binding.
- Streptavidin-HRP Conjugate (if biotinylated detection antibody is used):
- Enhances detection through enzymatic color change upon substrate addition.
- Substrate Solution (TMB):
- Tetramethylbenzidine, which reacts with HRP to produce a measurable colorimetric change.
Assay Procedure Summary
- Sample and Standard Addition: Samples and standards are added to the wells and incubated to allow TCR binding to the capture antibody.
- Wash: Unbound substances are washed away.
- Detection Antibody Addition: A secondary antibody, specific to another part of the TCR, is added and binds to the captured TCR.
- Wash: Removes unbound detection antibody.
- HRP-Streptavidin Conjugate Addition: (If using a biotinylated detection antibody) HRP-conjugated streptavidin is added, binding to the biotinylated detection antibody.
- Wash: Removes unbound conjugate.
Performance Characteristics
- Sensitivity: Detection limits in the picogram per milliliter (pg/mL) range.
- Precision: Intra-assay and inter-assay variability are generally low, with coefficients of variation (CV) typically below 10%.
- Linearity: Good linear response over the standard curve range.
- Recovery: Typically >80%, indicating the kit's robustness in different sample matrices.
Storage and Stability
- Storage: Components are usually stored at 2-8°C. The reconstituted standards and detection antibody solutions may require freezing at -20°C if not used immediately.
- Shelf Life: Generally 6-12 months under recommended storage conditions.
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