Triiodothyronine (T3) ELISA kits are designed to quantify the levels of T3 hormone in biological samples. T3 is a thyroid hormone critical for regulating metabolism, and its measurement is important in diagnosing thyroid disorders and assessing thyroid function.
Components
- Coated Microplate: A 96-well plate pre-coated with antibodies specific to T3. This allows for the capture of T3 from the samples.
- Standards: Recombinant or synthetic T3 with known concentrations to generate a standard curve for quantification.
- Detection Antibody: A secondary antibody specific to T3, typically conjugated to an enzyme such as horseradish peroxidase (HRP) for detection.
- Substrate Solution: Contains a substrate that reacts with HRP to produce a color change proportional to the amount of T3 present.
- Stop Solution: An acid solution used to stop the enzyme reaction and stabilize the color for measurement.
- Wash Buffer: Used to remove unbound substances and reduce background noise.
Procedure
- Sample Preparation: Dilute and add samples and standards to the wells of the microplate.
- Incubation: Allow T3 in the samples to bind to the antibodies on the plate.
- Detection: Add the detection antibody, which binds to the captured T3.
- Substrate Addition: Add substrate solution to develop a color proportional to T3 levels.
- Stopping Reaction: Add stop solution to halt the color development.
- Measurement: Measure optical density (OD) using a microplate reader to determine T3 concentration from the standard curve.
Technical Considerations
- Sensitivity: The lowest concentration of T3 that can be accurately detected by the assay.
- Specificity: The ability of the kit to specifically detect T3 without interference from other substances.
- Reproducibility: Consistency of results across different assays and conditions.
- Interferences: Potential effects of other substances in the sample on assay accuracy.
Applications
- Research: Studying T3's role in metabolism, thyroid function, and related disorders.
- Clinical Diagnostics: Assessing thyroid function and diagnosing conditions such as hyperthyroidism or hypothyroidism based on T3 levels.
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