Alkaline phosphatase (ALP) ELISA Kit – High-Quality Domestic ELISA Supplier
This ELISA kit is designed for the quantitative detection of mouse alkaline phosphatase (ALP) in various biological samples using a double-antibody sandwich method. The procedure involves coating a microplate with purified anti-mouse ALP antibodies, followed by incubation with the sample and HRP-conjugated secondary antibody. This forms a complex of antibody-antigen-enzyme-labeled antibody, which is then washed and developed using TMB substrate. Under the catalytic action of HRP, TMB turns blue and then changes to yellow when an acid is added. The intensity of the color is directly proportional to the concentration of ALP in the sample. The optical density (OD) is measured at 450 nm using a microplate reader, and the ALP concentration is determined by comparing the results to a standard curve.
Kit Components:
- 48-well configuration: 1×48 enzyme-labeled plate, 1 standard, 2 sealing films, 1 sealed bag, 1×48 reagents
- 96-well configuration: 1×96 enzyme-labeled plate, 1 standard, 2 sealing films, 1 sealed bag, 1×96 reagents
Storage Instructions:
All components should be stored at 2–8°C. The standard, enzyme reagent, sample diluent, developer A and B, and washing solution are provided in appropriate volumes.
Sample Preparation Guidelines:
1. **Serum**: Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes. Collect supernatant carefully; if precipitate forms, re-centrifuge.
2. **Plasma**: Use EDTA or sodium citrate as anticoagulant. Mix for 10–20 minutes, centrifuge, and collect supernatant. Re-centrifuge if necessary.
3. **Urine**: Collect in a sterile tube and centrifuge at 2000–3000 rpm for 20 minutes. Carefully collect supernatant.
4. **Cell Culture Supernatant**: For secreted proteins, collect supernatant after centrifugation. For intracellular components, lyse cells via freeze-thaw cycles and centrifuge again.
5. **Tissue Samples**: Weigh the tissue, add PBS (pH 7.4), homogenize, and centrifuge. Collect supernatant for testing. Store remaining samples at 2–8°C or -20°C.
6. **General Tips**: Process samples immediately after collection. If not tested right away, store at -20°C, avoiding repeated freeze-thaw cycles.
7. **Note**: Samples containing NaN3 cannot be used, as it inhibits HRP activity.
This kit ensures high sensitivity and specificity for ALP detection, making it ideal for research applications in immunology, toxicology, and clinical diagnostics.
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