TSH (Thyroid-Stimulating Hormone) ELISA kits are used to measure the concentration of TSH in biological samples, such as blood serum. TSH is a key hormone produced by the pituitary gland that regulates thyroid function. Accurate measurement of TSH levels is crucial for diagnosing thyroid disorders, including hypothyroidism and hyperthyroidism.
Components
- Coated Microplate: A 96-well plate pre-coated with antibodies specific to TSH. This allows TSH in the sample to be captured on the plate.
- Standards: Recombinant TSH with known concentrations used to generate a standard curve for quantifying TSH in samples.
- Detection Antibody: A secondary antibody specific to TSH, often conjugated to an enzyme like horseradish peroxidase (HRP) for detection.
- Substrate Solution: Contains a substrate that reacts with HRP to produce a color change, which is proportional to the amount of TSH.
- Stop Solution: An acid solution that stops the enzyme reaction, stabilizing the color for measurement.
- Wash Buffer: Used to remove unbound substances and reduce background interference.
Procedure
- Sample Preparation: Dilute and add samples and standards to the wells of the microplate.
- Incubation: Allow TSH in the samples to bind to the antibodies on the plate.
- Detection: Add the detection antibody, which binds to the captured TSH.
- Substrate Addition: Add substrate solution to develop a color proportional to TSH levels.
- Stopping Reaction: Add stop solution to halt color development.
- Measurement: Measure optical density (OD) using a microplate reader to determine TSH concentration from the standard curve.
Technical Considerations
- Sensitivity: The lowest concentration of TSH that can be accurately detected by the assay.
- Specificity: The ability to detect TSH specifically, without interference from other hormones or substances.
- Reproducibility: Consistency of results across different assays and sample conditions.
- Interferences: Potential effects of other substances in the sample that might impact assay accuracy.
Applications
- Research: Studying TSH’s role in thyroid regulation and related disorders.
- Clinical Diagnostics: Diagnosing and monitoring thyroid function disorders such as hypothyroidism and hyperthyroidism based on TSH levels.
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