Human Abnormal Prothrombin (APT) ELISA Technical Method Description – Guangrui Bio, a domestic leading supplier of high-quality ELISA kits
Sample Processing and Requirements:
1. **Serum**: Allow blood to clot at room temperature for 10–20 minutes. Centrifuge at approximately 2000–3000 rpm for 20 minutes. Carefully collect the supernatant. If precipitation occurs during storage, re-centrifuge before use.
2. **Plasma**: Use EDTA or sodium citrate as an anticoagulant based on sample requirements. Mix gently for 10–20 minutes, then centrifuge at 2000–3000 rpm for about 20 minutes. Carefully collect the supernatant. If a precipitate forms during storage, re-centrifuge before testing.
3. **Urine**: Collect using a sterile tube. Centrifuge at 2000–3000 rpm for 20 minutes. Carefully remove the supernatant. If a precipitate is observed, re-centrifuge. Similar procedures apply for pleural fluid, ascites, and cerebrospinal fluid.
4. **Cell Culture Supernatant**: For secreted components, collect in a sterile tube and centrifuge at 2000–3000 rpm for 20 minutes. Carefully collect the supernatant. For intracellular components, resuspend the cell pellet in PBS (pH 7.2–7.4), adjust the cell concentration to around 1 million cells/mL, and subject to repeated freeze-thaw cycles to lyse the cells. Centrifuge again at 2000–3000 rpm for 20 minutes. Carefully collect the supernatant. If precipitation occurs during storage, re-centrifuge.
5. **Tissue Specimen**: After excising the tissue, weigh it accurately. Add a suitable volume of PBS (pH 7.4). Immediately freeze in liquid nitrogen and store at -80°C. When thawing, ensure the specimen remains between 2–8°C. Add more PBS, homogenize manually or with a homogenizer, and centrifuge at 2000–3000 rpm for 20 minutes. Carefully collect the supernatant. Use a portion immediately and store the rest for future use.
6. **General Sample Handling**: Process samples as soon as possible after collection. Follow relevant literature for extraction procedures. Perform the ELISA test promptly after sample preparation. If testing cannot be done immediately, store the samples at -20°C. Avoid repeated freezing and thawing to prevent degradation.
7. **Note**: Samples containing sodium azide (NaN₃) should not be used, as NaN₃ can inhibit horseradish peroxidase (HRP) activity, which is critical for the ELISA detection system.
By following these detailed protocols, you can ensure accurate and reliable results when performing Human Abnormal Prothrombin (APT) ELISA assays using Guangrui Bio’s high-performance ELISA kits.
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