Mouse interleukin 18 (IL-18) elisa kit instruction manual

**Mouse IL-18 (Interleukin-18) ELISA Kit – Instructions for Use** This ELISA kit is designed for the quantitative determination of mouse Interleukin-18 (IL-18) in serum, plasma, urine, cell culture supernatants, and tissue homogenates. The kit uses a sandwich ELISA method, which ensures high specificity and sensitivity for IL-18 detection. **Kit Specifications:** - **Configuration:** 48-well or 96-well format - **Standard Dilution:** 1.5 mL × 1 vial - **Enzyme Standard Reagent:** 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well) - **Storage Conditions:** 2–8°C - **Shelf Life:** 6 months from date of manufacture **Kit Components:** - Sealing Film: 2 pieces (48-well), 2 pieces (96-well) - Standard: 0.5 mL × 1 vial, 2700 ng/L - Enzyme Standard: 1×48, 1×96 - Sample Diluent: 3 mL × 1 vial, 6 mL × 1 vial - Developer A: 3 mL × 1 vial, 6 mL × 1 vial - Chromogen B: 3 mL × 1 vial, 6 mL × 1 vial - Wash Solution: 3 mL × 1 vial, 6 mL × 1 vial - Concentrated Wash Solution: 20 mL × 20 times, 20 mL × 30 times **Principle of Operation:** The ELISA kit employs a double-antibody sandwich method. The microplate is pre-coated with anti-mouse IL-18 monoclonal antibody. After adding the sample, IL-18 binds to the coated antibody. HRP-conjugated secondary antibody then binds to the complex. TMB substrate is added, and the color develops in proportion to the IL-18 concentration. The reaction is stopped with an acidic solution, and the absorbance at 450 nm is measured. A standard curve is plotted using known concentrations, and the sample concentration is determined accordingly. **Sample Preparation Guidelines:** - **Serum/Plasma:** Centrifuge at 2000–3000 rpm for 20 minutes after clotting or anticoagulant addition. - **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes. - **Cell Culture Supernatant:** Centrifuge after collection; for intracellular components, lyse cells via freeze-thaw cycles. - **Tissue Homogenate:** Homogenize in PBS, centrifuge, and collect supernatant. **Operation Steps:** 1. Prepare standard dilutions (1800, 1200, 600, 300, 150 ng/L). 2. Add sample diluent and test samples (5x dilution). 3. Incubate at 37°C for 30 minutes. 4. Wash plate 5 times with diluted wash solution. 5. Add enzyme-labeled reagent. 6. Incubate again for 30 minutes. 7. Add TMB substrate and incubate for 15 minutes. 8. Stop the reaction with stop solution. 9. Measure OD at 450 nm within 15 minutes. **Notes and Recommendations:** - Allow the kit to equilibrate to room temperature before use. - Avoid cross-contamination by using a new sealing film for each experiment. - Always prepare a standard curve with duplicate wells. - If sample OD exceeds that of the highest standard, perform a preliminary dilution. - Handle all reagents with care and follow biosafety protocols. - Store unused components at 2–8°C. **Performance:** - Linear range: 0.2 IU/L – 6 IU/L - Correlation coefficient (R²): ≥ 0.95 - Intra-batch and inter-batch variation: <9% and <11%, respectively **Technical Support:** Our team offers free technical assistance during working hours. For more information or to request sample testing, please contact us directly. **Note:** This kit is for research use only. Do not use if the contents are expired or damaged.

School Chairs

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