KRT20 (Keratin 20) ELISA kits are designed to measure the levels of Keratin 20 protein in biological samples. This is valuable for research and clinical diagnostics, particularly in studying epithelial cell behavior and various cancers.
Components
- Coated Microplate: A 96-well plate pre-coated with antibodies specific to KRT20. This allows for the capture of KRT20 from the samples.
- Standards: Recombinant KRT20 proteins with known concentrations, used to generate a standard curve for quantification.
- Detection Antibody: A secondary antibody specific to KRT20, 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 KRT20 present.
- Stop Solution: An acid solution used to stop the enzyme reaction and stabilize the color for measurement.
- Wash Buffer: Used to wash away unbound proteins and reduce background noise.
Procedure
- Sample Preparation: Dilute and add samples and standards to the wells of the microplate.
- Incubation: Allow KRT20 in the samples to bind to the antibodies on the plate.
- Detection: Add the detection antibody, which binds to the captured KRT20.
- Substrate Addition: Add substrate solution to develop a color proportional to KRT20 levels.
- Stopping Reaction: Add stop solution to halt the color development.
- Measurement: Measure optical density (OD) using a microplate reader to determine KRT20 concentration from the standard curve.
Technical Considerations
- Sensitivity: The minimum detectable concentration of KRT20.
- Specificity: The ability of the kit to specifically detect KRT20 without interference from other proteins.
- Reproducibility: Consistency of assay results across different tests and conditions.
- Interferences: Potential effects of other substances in the sample on assay accuracy.
Applications
- Research: Investigating the role of KRT20 in epithelial cells and various cancers, such as gastrointestinal tumors.
- Clinical Diagnostics: Monitoring KRT20 levels for diagnostic and prognostic purposes in conditions associated with epithelial cell abnormalities.
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