Monkey B virus IgG (BV IgG) ELISA kit instruction manual

**Monkey B Virus IgG (BV IgG) ELISA Kit Instructions for Use** This kit is intended for research use only and is designed to detect the presence of Monkey B Virus IgG (BV IgG) in serum, plasma, and other biological fluids from monkeys. The test is based on a double-antibody sandwich ELISA method, which allows for highly specific and sensitive detection of BV IgG. **Principle of the Test** The microplate is pre-coated with a purified monoclonal antibody against BV IgG. When the sample is added, any BV IgG present will bind to the immobilized antibody. A horseradish peroxidase (HRP)-labeled secondary antibody then binds to the captured BV IgG, forming an immune complex. After washing to remove unbound components, a TMB substrate is added. Under the catalytic action of HRP, TMB changes color from blue to yellow. The reaction is stopped by adding an acidic solution, and the absorbance at 450 nm is measured using a microplate reader. Results are compared to a cutoff value to determine whether the sample is positive or negative for BV IgG. **Kit Components** The kit includes all necessary reagents for 48 or 96 tests, depending on the configuration. It contains: - Microtiter plates coated with anti-BV IgG antibodies - Negative and positive controls - Enzyme-labeled anti-BV IgG antibody - TMB substrate solution - Wash buffer - Sample diluent - Stop solution - Concentrated wash solution (for multiple uses) **Storage Conditions** All components should be stored at 2–8°C. The kit should be kept sealed and protected from light until use. **Sample Preparation and Handling** Proper handling of samples is essential for accurate results. Here are guidelines for different types of specimens: 1. **Serum**: Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes. Collect the supernatant carefully. 2. **Plasma**: Use EDTA or sodium citrate as anticoagulant. Mix well and centrifuge as above. 3. **Urine**: Collect in sterile tubes and centrifuge at 2000–3000 rpm for 20 minutes. 4. **Cell Culture Supernatant**: Centrifuge after collection. For intracellular components, lyse cells by repeated freezing and thawing before centrifugation. 5. **Tissue Samples**: Homogenize in PBS, centrifuge, and collect the supernatant. Store at 2–8°C if not used immediately. 6. **General Notes**: Avoid repeated freeze-thaw cycles. Do not use samples containing NaN3, as it may interfere with the HRP enzyme. **Procedure** 1. Label the wells and set up duplicate negative and positive controls, along with a blank control. 2. Add 50 µL of negative/positive control and 50 µL of sample diluent, followed by 10 µL of the sample into each well. 3. Incubate at 37°C for 30 minutes. 4. Wash the plate five times with diluted wash buffer. 5. Add 50 µL of enzyme-labeled reagent to each well (except the blank). 6. Incubate again at 37°C for 30 minutes. 7. Wash again. 8. Add 50 µL of developer A and 50 µL of developer B, incubate for 15 minutes at 37°C. 9. Stop the reaction with 50 µL of stop solution. 10. Measure OD at 450 nm within 15 minutes. **Interpretation of Results** - **Positive Control Mean ≥ 1.00** - **Negative Control Mean ≤ 0.10** - **Cutoff Value = Negative Control Mean + 0.15** If the sample OD is below the cutoff, the result is considered negative; otherwise, it is positive. **Note:** Always follow safety protocols when handling biological materials. Ensure proper disposal of waste according to local regulations. **Download the Full Manual** [Click here to download the complete instruction manual](#). For more information or to purchase this kit, visit the [Shanghai Kamaishu Laboratory Research Reagents Procurement Network](#). *Stay updated with the latest in laboratory supplies and research tools.*

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