Human CXCL16 ELISA Kit

**Human CXCL16 ELISA Kit – For the Quantitative In Vitro Determination of Human CXC-chemokine Ligand 16 Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids** **For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.** This ELISA kit is designed for the accurate and reliable quantification of human CXCL16 in various biological samples. The assay is based on a sandwich immunoassay principle, utilizing specific antibodies to capture and detect CXCL16 in the sample. A standard curve is generated using a series of calibration standards, allowing for precise determination of CXCL16 concentrations in unknown samples. **Intended Use:** The Human CXCL16 ELISA Kit is intended for research purposes only and is not suitable for diagnostic or clinical use. It provides a sensitive and specific method for measuring CXCL16 levels in biological fluids such as serum, plasma, cerebrospinal fluid, tissue homogenates, and other body fluids. **Principle of the Assay:** The kit includes a set of calibration standards that are run alongside the samples. After incubation with the HRP-conjugated antibody, the colorimetric reaction is stopped, and the optical density (OD) is measured at 450 nm. The concentration of CXCL16 in the samples is then calculated by comparing their OD values to the standard curve. **Sample Collection and Storage:** - **Serum:** Collect in serum separator tubes, allow clotting for 2 hours at room temperature or overnight at 4°C, then centrifuge at ~2000×g for 20 minutes. Store at -20°C. Avoid repeated freeze-thaw cycles. - **Plasma, Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids:** Centrifuge to remove particulates, and either assay immediately or aliquot and store at -20°C. Ensure no hemolysis or contamination occurs. **Materials Required but Not Supplied:** - 37°C Incubator - Microplate Reader (measuring absorbance at 450 nm) - Precision Pipettes, Disposable Tips, and Absorbent Paper - Distilled or Deionized Water **Reagents Provided (Stored at 2–8°C):** - Microtiter Strip Plate (12×8 or 12×4 strips) - Standards (6 vials, 0.5 ml/vial) - Sample Diluent (6.0 ml or 3.0 ml) - HRP-Conjugate Reagent (10.0 ml or 5.0 ml) - 20X Wash Solution (25 ml or 15 ml) - Chromogen Solution A (6.0 ml or 3.0 ml) - Chromogen Solution B (6.0 ml or 3.0 ml) - Stop Solution (6.0 ml or 3.0 ml) - Closure Plate Membrane (2 units) - User Manual (1 copy) - Sealed Bags (1 unit) **Standard Concentrations:** 20, 10, 5, 2.5, 1.25, 0.625 ng/ml **Preparation and Storage Notes:** - All reagents should be brought to room temperature (20–25°C) before use. - Do not use water baths for thawing. - Do not use reagents beyond their expiration date. - Only use deionized or distilled water for dilutions. - Keep microtiter plates in sealed bags until needed. Unused strips should be stored with desiccant at 2–8°C. - Use fresh pipette tips for each transfer to avoid cross-contamination. **Safety and Handling:** - Handle all biological samples with care. Wear gloves during the procedure. - Disposal of waste must include a minimum of 30 minutes inactivation time before disposal. - Substrate Solution A may change color if contaminated; do not use if it turns blue. - Substrate B contains 20% acetone—keep away from heat and flame. **Assay Procedure:** 1. Prepare all reagents before starting. Run standards and samples in duplicate. 2. Add 50 µl of standard or sample to appropriate wells. Blank well receives no addition. 3. Add 100 µl of HRP-conjugate reagent to all wells except blank. Cover and incubate for 60 minutes at 37°C. 4. Wash the plate 4 times using manual or automated washing. 5. Add 50 µl of Chromogen A and 50 µl of Chromogen B. Incubate for 15 minutes at 37°C, protected from light. 6. Add 50 µl of Stop Solution. The color should turn yellow. If green or uneven, gently mix. 7. Measure OD at 450 nm. Generate a standard curve and calculate sample concentrations. **Performance Characteristics:** - Intra-assay CV < 15% - Inter-assay CV < 15% - Sensitivity: < 0.1 ng/ml - Dynamic Range: 0.625 – 20 ng/ml - Cross-reactivity: No significant cross-reactivity with other chemokines observed. **Storage Conditions:** - Store unused kit at 2–8°C for frequent use, or -20°C for long-term storage (up to 6 months). **Note:** Always read the entire protocol carefully before beginning the assay. Each user should generate their own standard curve for best results.

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