CXCL8, also known as Interleukin-8 (IL-8), is a chemokine involved in the recruitment and activation of neutrophils during inflammation. CXCL8 ELISA kits are used to quantitatively measure CXCL8 levels in biological samples such as serum, plasma, urine, or cell culture supernatants.
Overview of CXCL8 ELISA Kits
- Purpose: To detect and quantify CXCL8/IL-8 in various biological fluids.
- Assay Principle: Usually a sandwich ELISA. The CXCL8 protein in the sample is captured by a specific antibody, followed by detection using a secondary antibody conjugated to an enzyme (typically HRP). A substrate reaction generates a measurable color change, correlating with CXCL8 concentration.
- Sensitivity: High sensitivity, allowing detection of CXCL8 in the low picogram per milliliter (pg/mL) range.
- Specificity: Highly specific to CXCL8, with minimal cross-reactivity to other chemokines or cytokines.
- Sample Volume: Typically requires 50-100 µL of sample per well.
Content of a Standard CXCL8 ELISA Kit
- Microtiter Plate: Pre-coated with a monoclonal or polyclonal anti-CXCL8 antibody.
- Standards: Recombinant CXCL8 at various concentrations to generate a standard curve.
- Detection Antibody: Biotinylated or enzyme-conjugated anti-CXCL8 antibody.
- Enzyme Conjugate: Typically streptavidin-HRP for signal amplification.
- Substrate Solution: Commonly TMB (3,3',5,5'-Tetramethylbenzidine), which produces a color change upon reaction.
- Stop Solution: Usually a sulfuric acid solution to halt the enzymatic reaction and stabilize the color.
Key Steps in the Assay Procedure
- Sample/Standard Addition: Standards and samples are added to the wells coated with anti-CXCL8 antibodies.
- Incubation: The plate is incubated, allowing CXCL8 to bind to the capture antibody.
- Washing: Wells are washed to remove unbound components.
- Detection Antibody Addition: The detection antibody is added and binds to the captured CXCL8.
- Washing: Unbound detection antibody is washed away.
- Enzyme Conjugate Addition: The enzyme conjugate is added, which binds to the detection antibody.
Applications
- Research: Studying the role of CXCL8 in inflammation, immune response, cancer, and other diseases.
- Clinical Diagnostics: Measuring CXCL8 levels to evaluate inflammatory conditions, infections, and cancer prognosis.
CXCL8 ELISA kits are critical for investigating the chemokine’s role in various biological processes and for monitoring inflammatory responses in both research and clinical settings.
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