Chicken sIgA ELISA Kit

**Chicken sIgA ELISA Kit – For the Quantitative In Vitro Determination of Chicken Secretory Immunoglobulin A 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 quantitative determination of secretory immunoglobulin A (sIgA) in various biological samples. The test is based on the enzyme-linked immunosorbent assay (ELISA) principle, using a competitive binding format. The Stop Solution changes the color from blue to yellow, and the intensity of the color is directly proportional to the concentration of sIgA in the sample. To measure sIgA levels, a set of calibration standards is included. These standards are run alongside the samples to generate a standard curve, which allows for accurate quantification of sIgA in unknown samples by comparing their optical density (OD) values at 450 nm to the standard curve. --- **Sample Collection and Storage** - **Serum**: Collect using a serum separator tube. Allow samples to clot for 2 hours at room temperature or overnight at 4°C. Centrifuge at 2000×g for 20 minutes. Remove serum and assay immediately, or aliquot and store at -20°C. Avoid repeated freeze-thaw cycles. - **Plasma**: Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g for 30 minutes at 2–8°C. Store at -20°C. - **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids**: Centrifuge to remove particulates. Assay immediately or aliquot and store at -20°C. Ensure no hemolysis or granules are present in the samples. --- **Materials Required but Not Supplied** 1. Incubator at 37°C 2. Microplate reader capable of measuring absorbance at 450 nm 3. Precision pipettes, disposable tips, and absorbent paper 4. Distilled or deionized water --- **Reagents Provided** - Microtiter Strip Plate (12×8 or 12×4 strips) - Standards (6 vials, 0.5 mL/vial) – concentrations: 8, 4, 2, 1, 0.5, 0.25 ng/mL - 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 pieces) - User Manual (1 copy) - Sealed Bags (1 bag) --- **Important Notes** - All reagents should be stored at 2–8°C. Check expiration date on the label before use. - Do not use reagents beyond their expiration date. - Only use deionized or distilled water for dilutions. - Allow all reagents and samples to reach room temperature (20–25°C) before use. - Avoid thawing samples or reagents in a water bath. - Keep microtiter plate in its sealed bag until ready for use. Unused strips should be stored with desiccant. - Use fresh disposable pipette tips for each transfer to prevent cross-contamination. - Dispose of all samples and reagents following proper biohazard protocols. - Liquid waste should be treated with sodium hypochlorite (final concentration 1.0%) and left for at least 30 minutes before disposal. - Chromogen Solution B contains 20% acetone; keep away from heat or flame. - Substrate solutions are sensitive to contamination; avoid exposure to light. --- **Assay Procedure** 1. Prepare all reagents and ensure they are at room temperature. 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 the blank. Cover with adhesive strip and incubate for 60 minutes at 37°C. 4. Wash the plate 4 times using either manual or automated washing methods. 5. Add 50 µL of Chromogen A and 50 µL of Chromogen B to each well. Mix gently and incubate for 15 minutes at 37°C, protecting from light. 6. Add 50 µL of Stop Solution to each well. Read OD at 450 nm using a microplate reader. 7. Generate a standard curve by plotting average OD values against standard concentrations. 8. Calculate the mean OD for each standard and sample, subtracting the blank value. 9. Determine sIgA concentration by comparing sample OD to the standard curve. --- **Performance Characteristics** - **Intra-assay CV (%) and Inter-assay CV (%)**: <15% - **Assay Range**: 0.25 ng/mL – 8 ng/mL - **Sensitivity**: <0.1 ng/mL - **Cross-reactivity**: No significant cross-reactivity observed with other immunoglobulins. - **Storage**: 2–8°C (for frequent use); 6 months at -20°C. This kit is intended solely for research purposes and must not be used in diagnostic procedures. Always follow good laboratory practices and safety guidelines when handling biological materials.

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