Human papillomavirus L1 antibody (HPVL1 Ab) elisa kit manufacturer description

Human papillomavirus L1 antibody (HPV L1 Ab) sample preparation and handling instructions: 1. **Serum**: Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for approximately 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, depending on the test requirements. Mix gently for 10–20 minutes, then centrifuge at 2000–3000 rpm for about 20 minutes. Collect the supernatant carefully and re-centrifuge if needed. 3. **Urine**: Collect in a sterile tube and centrifuge at 2000–3000 rpm for 20 minutes. Carefully remove the supernatant. If precipitates form, re-centrifuge. Similar procedures apply to 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. For intracellular components, dilute the cell suspension in PBS (pH 7.2–7.4) to reach ~1 million cells/mL. Freeze-thaw repeatedly to lyse cells, then centrifuge again to collect the supernatant. Store and re-centrifuge if necessary. 5. **Tissue specimens**: Weigh the tissue after cutting, add PBS (pH 7.4), freeze quickly in liquid nitrogen, and store at 2–8°C after thawing. Homogenize with a mortar or homogenizer, centrifuge at 2000–3000 rpm for 20 minutes, and collect the supernatant. One portion is tested immediately; the rest is frozen for future use. 6. **General handling**: Process samples as soon as possible after collection, following standard protocols. If not used immediately, store at -20°C. Avoid repeated freezing and thawing to maintain integrity. 7. **Avoidance of NaN3**: Samples containing sodium azide cannot be tested, as it inhibits horseradish peroxidase (HRP) activity. **Procedure steps:** 1. **Standard preparation**: Place 10 standard wells on the microplate. Add 100 μL of standard to the first and second wells, then mix 50 μL with standard diluent. Transfer 100 μL from each to the next two wells, followed by 50 μL diluent. Continue this serial dilution to create concentrations of 9 μg/L, 6 μg/L, 3 μg/L, 1.5 μg/L, and 0.75 μg/L. 2. **Sample addition**: Set up blank wells (without sample or enzyme reagent). Add 40 μL of sample diluent to each test well, then 10 μL of the sample (final 5× dilution). Add to the bottom of the well, avoid touching the walls, and mix gently. 3. **Incubation**: Seal the plate and incubate at 37°C for 30 minutes. 4. **Washing solution**: Dilute 30× (or 20× for 48T) concentrated wash buffer with distilled water. 5. **Washing**: Remove the seal, discard the liquid, and fill each well with washing solution. Let stand for 30 seconds, discard, repeat 5 times, and pat dry. 6. **Enzyme conjugate**: Add 50 μL of enzyme-labeled reagent to each well except blanks. 7. **Second incubation**: Repeat step 3. 8. **Second washing**: Follow step 5. 9. **Color development**: Add 50 μL of color reagent A and B, mix gently, and incubate at 37°C for 15 minutes away from light. 10. **Stop reaction**: Add 50 μL of stop solution to each well to halt the reaction (color changes from blue to yellow). 11. **Measurement**: Read OD values at 450 nm within 15 minutes after stopping the reaction. Ensure the blank well is zeroed before measuring. This detailed procedure ensures accurate and reliable detection of HPV L1 antibodies while maintaining sample integrity and minimizing contamination risks.

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