Is the sample processing required by the ELISA kit required and clear? -Huaqiang Electronic Network

Recently, a first-year student from Nantong University purchased our ELISA kit and reached out to us for guidance on sample preparation and requirements. To help more people better understand the process, we’ve put together this detailed guide: **Serum Preparation:** After collecting blood, allow it to clot at room temperature for 10–20 minutes. Then centrifuge at 2000–3000 rpm for about 20 minutes. Carefully collect the supernatant. If precipitation occurs during storage, re-centrifuge before use. Always use endotoxin-free and pyrogen-free tubes to avoid contamination. **Plasma Handling:** Use EDTA or sodium citrate as an anticoagulant depending on your needs. Mix gently for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes. Carefully collect the supernatant. If precipitates form during storage, re-centrifuge before testing. **Urine Sample:** Collect urine in a sterile tube and centrifuge at 2000–3000 rpm for 20 minutes. Carefully remove the supernatant. If precipitation occurs, re-centrifuge. Similar procedures apply to pleural fluid, ascites, and cerebrospinal fluid. **Cell Culture Supernatant:** For secreted components, collect the supernatant directly after centrifugation (2000–3000 rpm, 20 min). For intracellular components, resuspend the cell pellet in PBS (pH 7.2–7.4) to a concentration of ~1 million cells/mL. Freeze and thaw repeatedly to lyse the cells, then centrifuge again and collect the supernatant. Re-centrifuge if needed. **Tissue Specimens:** Weigh the tissue sample after cutting, then add PBS (pH 7.4). Rapidly freeze in liquid nitrogen and store at -80°C. When thawing, keep the sample at 2–8°C. Add PBS, homogenize using a pestle or homogenizer, and centrifuge at 2000–3000 rpm for 20 minutes. Use one portion immediately and freeze the rest for future use. **Important Notes:** Process samples as soon as possible after collection, following standard protocols. If testing is delayed, store at -20°C but avoid repeated freezing and thawing. Sodium azide (NaN3) should not be present in samples, as it can inhibit horseradish peroxidase (HRP), which is commonly used in ELISA assays. If you have any questions or need further assistance, feel free to reach out. We’re here to help!

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