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

Human papillomavirus L1 antibody (HPV L1 Ab) sample preparation and handling instructions: 1. **Serum**: After blood is left at room temperature for 10–20 minutes to clot, 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 anticoagulant based on specimen requirements. Mix for 10–20 minutes, then centrifuge at 2000–3000 rpm for about 20 minutes. Collect the supernatant carefully. If precipitate forms during storage, re-centrifuge. 3. **Urine**: Collect in a sterile tube and centrifuge at 2000–3000 rpm for 20 minutes. Carefully remove the supernatant. If precipitate appears during storage, 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. For intracellular components, dilute cell suspension with PBS (pH 7.2–7.4) to achieve ~1 million cells/mL. Freeze-thaw repeatedly to lyse cells, then centrifuge again. Collect the supernatant carefully and re-centrifuge if needed. 5. **Tissue samples**: Weigh the tissue after cutting, add PBS (pH 7.4), and quickly freeze in liquid nitrogen. Store at 2–8°C after thawing. Add PBS, homogenize manually or using a homogenizer, then centrifuge at 2000–3000 rpm for 20 minutes. Collect the supernatant, test a portion immediately, and store the rest. 6. **General handling**: Process specimens as soon as possible after collection. Follow established protocols for extraction. Perform the assay promptly. If not done immediately, store at -20°C but avoid repeated freezing and thawing. 7. **Avoid NaN3**: Samples containing sodium azide cannot be tested, as it inhibits horseradish peroxidase (HRP). **Procedure Steps**: 1. **Standard preparation**: Place 10 standard wells on the microplate. Add 100 μL of standard to the first and second wells, mix with 50 μL diluent. Transfer 100 μL from each to the next wells, adding 50 μL diluent each time. Dilute serially to obtain concentrations of 9, 6, 3, 1.5, and 0.75 μg/L. 2. **Sample addition**: Set up blank wells (no sample or enzyme reagent). Add 40 μL sample diluent to each test well, followed by 10 μL of the sample (final 5× dilution). Avoid touching the well walls and gently mix. 3. **Incubation**: Seal the plate and incubate at 37°C for 30 minutes. 4. **Washing solution**: Dilute 30× (for 20T) or 20× (for 48T) concentrated wash solution with distilled water. 5. **Washing**: Remove the seal, discard liquid, and fill each well with wash solution. Incubate for 30 seconds, discard, repeat 5 times, and pat dry. 6. **Enzyme labeling**: Add 50 μL of enzyme-labeled reagent to each well except blanks. 7. **Incubation**: Repeat step 3. 8. **Washing**: Repeat 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 stop solution to each well. The color changes from blue to yellow. 11. **Measurement**: Read OD values at 450 nm within 15 minutes of adding the stop solution. Ensure the blank well is set to zero before measuring. This detailed procedure ensures accurate and consistent results when detecting HPV L1 antibodies. Always follow safety protocols and maintain proper storage conditions to preserve sample integrity.

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