Human EB virus detection plate (EB) ELISA test kit instruction manual

**Human Epstein-Barr Virus Detection Plate (EB) Enzyme-Linked Immunosorbent Assay (ELISA) Kit Instruction Manual** This reagent is intended for research purposes only. The Human EB Virus Detection Plate (EB) ELISA Kit is designed to detect the presence of EB virus in various human biological samples, including serum, plasma, cell culture supernatant, and other related liquid specimens. It supports the identification and hematological evaluation of individuals infected with EB virus. **Principle of the Test** The kit utilizes a double-antibody sandwich ELISA method to quantify the EB virus detection plate (EB) in the sample. A microplate pre-coated with purified EB-specific antibodies is used as a solid-phase carrier. These antibodies bind to EB antigens present in the sample. After washing away unbound components, an HRP-labeled secondary antibody is added to form a complex. Following another wash, TMB substrate is introduced, which changes color under the catalytic action of HRP. The reaction is stopped by adding an acidic solution, resulting in a yellow color. The optical density (OD) at 450 nm is measured using a microplate reader, and the results are compared against a cutoff value to determine the presence or absence of EB in the specimen. **Kit Components** - 48-well configuration: 1 × 48 enzyme-labeled microplate, 2 sealing films, 1 negative control, 1 positive control, 1 standard reagent, 1 sample diluent, 1 developer A, 1 developer B, 1 stop solution, 1 concentrated washing solution (20×, 20 ml) - 96-well configuration: 1 × 96 enzyme-labeled microplate, 2 sealing films, 1 negative control, 1 positive control, 1 standard reagent, 1 sample diluent, 1 developer A, 1 developer B, 1 stop solution, 1 concentrated washing solution (20×, 30 ml) All components should be stored at 2–8°C. **Sample Preparation and Handling** 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; if precipitate forms, re-centrifuge before use. 2. **Plasma**: Use EDTA or sodium citrate as anticoagulant. Mix well and centrifuge at 2000–3000 rpm for 20 minutes. Carefully collect the supernatant. 3. **Urine**: Collect using a sterile container and centrifuge at 2000–3000 rpm for 20 minutes. Remove the supernatant. 4. **Cell Culture Supernatant**: Centrifuge at 2000–3000 rpm for 20 minutes. For intracellular components, lyse cells by repeated freeze-thaw cycles and centrifuge again. 5. **Tissue Specimens**: Weigh the tissue, add PBS (pH 7.4), homogenize, and centrifuge. Store at 2–8°C after thawing. 6. **Storage**: Process samples immediately after collection. If not tested right away, store at -20°C, avoiding repeated freeze-thaw cycles. Do not use samples containing NaN₃, as it may inhibit HRP activity. **Procedure** 1. Label all wells. Set up 2 negative control, 2 positive control, and 1 blank control per plate. 2. Add 50 μl of negative/positive control, 40 μl of sample diluent, and 10 μl of sample into respective wells. Gently mix. 3. Incubate at 37°C for 30 minutes. 4. Prepare washing solution by diluting the concentrated washing solution with distilled water. 5. Wash the plate 5 times, each time for 30 seconds. 6. Add 50 μl of HRP-labeled antibody to each well except the blank. 7. Incubate again at 37°C for 30 minutes. 8. Wash the plate again. 9. Add 50 μl of developer A followed by 50 μl of developer B. Incubate at 37°C for 15 minutes. 10. Stop the reaction by adding 50 μl of stop solution. 11. Measure OD at 450 nm within 15 minutes. **Result Interpretation** - **Validity Check**: Positive control OD ≥ 1.00, Negative control OD ≤ 0.10. - **Cutoff Value**: Cutoff = Average of negative control + 0.15 - **Negative Result**: Sample OD < Cutoff - **Positive Result**: Sample OD ≥ Cutoff **Note**: This manual is provided for reference. Always follow the manufacturer’s instructions and ensure proper training before performing the assay. **Download**: [Human EB Virus Detection Plate (EB) ELISA Test Kit Instruction Manual](#) **Author**: [Shanghai Kamaishu Laboratory Research Reagents Procurement Network](#) **Keywords**: EB, ELISA, Viral Detection, Diagnostic Kit, Research Use Only **Copyright Notice**: All content on this site is protected by copyright. Unauthorized reproduction or distribution is prohibited. Please cite "Source: China Education Equipment Purchasing Network" when using any content from this site. **QR Code**: [Scan to Access More Information](#) **Official WeChat**: Follow us for the latest updates on education equipment and research tools. **About Us**: China Education Equipment Purchasing Network – Your trusted source for laboratory reagents and equipment. Stay informed and connected.

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