**Goat IgA ELISA Kit – For the Quantitative In Vitro Determination of Goat Immunoglobulin A in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Biological Fluids** *For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.* Before using this product, please read this entire package insert carefully. This ELISA kit is designed for research purposes only and should not be used in clinical diagnostic settings. **Intended Use & Test Principle** The Goat IgA ELISA Kit is intended for laboratory research use only and is not suitable for diagnostic or therapeutic applications. The test is based on the enzyme-linked immunosorbent assay (ELISA) principle. The concentration of IgA in the sample is determined by comparing the optical density (OD) of the sample to a standard curve generated from known concentrations of IgA standards. A stop solution is used to terminate the reaction, changing the color from blue to yellow, allowing quantification of IgA levels based on the intensity of the color. **Sample Collection and Storage** - **Serum**: Use a serum separator tube. Allow samples to clot for 2 hours at room temperature or overnight at 4°C before centrifuging at 2000×g for 20 minutes. 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, 2–8°C. Store at -20°C. Avoid repeated freezing. - **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids**: Centrifuge to remove particulates. Assay immediately or store at -20°C. Ensure no hemolysis or granules are present. **Materials Required but Not Supplied** 1. Incubator set at 37°C 2. Microplate reader capable of measuring absorbance at 450 nm 3. Pipettes, disposable tips, and absorbent paper 4. Distilled or deionized water **Reagents Provided (Stored at 2–8°C)** - MicroElisa Strip Plate: 12×8 strips / 12×4 strips - Standard (6 vials): 0.5 ml/vial - Sample Diluent: 6.0 ml / 3.0 ml - HRP-Conjugate Reagent: 10.0 ml / 5.0 ml - 20X Wash Solution: 25 ml / 15 ml - Chromogen Solution A: 6.0 ml / 3.0 ml - Chromogen Solution B: 6.0 ml / 3.0 ml - Stop Solution: 6.0 ml / 3.0 ml - Closure Plate Membrane: 2 / 2 - User Manual: 1 / 1 - Sealed Bags: 1 / 1 **Standard Concentrations**: 48, 24, 12, 6, 3, 1.5 μg/mL If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay. **Important Notes** - Do not thaw samples or reagents in a water bath. - Do not use any kit components past their expiration date. - Always use deionized or distilled water for dilutions. - Keep microtiter plates in sealed bags until use. Unused strips should be stored with desiccant at 2–8°C. - Use fresh pipette tips for each transfer to prevent cross-contamination. - Disposable knives must not be used in the assay due to potential risk of infectious agents. - All samples should be treated as potentially infectious. Follow biosafety guidelines. - Dispose of all samples and waste according to local regulations. - Liquid waste: Add 1% sodium hypochlorite and let stand for 30 minutes before disposal. - Substrate solutions are sensitive; avoid contamination. If the solution appears bluish, discard it. - Chromogen B contains 20% acetone; keep away from heat and flame. - Bring all reagents to room temperature (20–25°C) before use. **Reagent Preparation and Storage** - **Wash Solution (1X)**: Dilute 1 volume of 20X Wash Solution with 19 volumes of deionized or distilled water. Store at 2–8°C for up to one month. **Assay Procedure** 1. Prepare all reagents before starting. Run standards and samples in duplicate. 2. Add 50 µl of standard or sample to the appropriate wells. Blank well receives nothing. 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. - **Manual Washing**: Aspirate contents, fill with 1X Wash Solution, aspirate again. Repeat 4 times. Dry by tapping the plate. - **Automated Washing**: Aspirate and wash four times with 350 µL/well. Dry by tapping. 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 within 15 minutes. **Calculation** - Plot average OD values (450 nm) against standard concentrations. - Subtract blank OD from all readings before interpreting results. - Construct the standard curve using graph paper or software. - At the point of intersection, draw a vertical line to the X-axis to determine IgA concentration. - Intra-assay CV: <15%, Inter-assay range: 1.5–48 µg/mL. - Sensitivity: <1.0 µg/mL. - No significant cross-reactivity or interference observed. **Storage** - Store at 2–8°C for frequent use. - For long-term storage, keep at -20°C for up to 6 months. **Safety and Handling** Handle all materials with care. Follow lab safety protocols and dispose of waste properly. **Note**: This kit is not approved for diagnostic use. Always follow the manufacturer’s instructions and maintain proper documentation.

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