**Alkaline Phosphatase (ALP) ELISA Kit – Technical Specifications**
Guangrui Bio, a trusted domestic supplier of high-quality ELISA kits, offers this reliable and accurate Alkaline Phosphatase (ALP) ELISA kit for the quantitative detection of mouse ALP in various biological samples. This kit employs the double-antibody sandwich ELISA method, ensuring high specificity and sensitivity.
The procedure begins with coating a microtiter plate with purified anti-mouse ALP antibodies to create a solid-phase antibody. After adding the sample, the target ALP binds to the immobilized antibody. A horseradish peroxidase (HRP)-labeled anti-ALP antibody is then introduced, forming a complex of antibody-antigen-enzyme-labeled antibody. Following thorough washing, TMB substrate is added, leading to a color change. Under HRP catalysis, TMB turns blue and then yellow upon acid termination. The intensity of the color is directly proportional to the concentration of ALP in the sample.
The absorbance 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 seal film, 2 sealed bags, 1 standard, 1 standard diluent, 1 sample diluent, 1 developer A, 1 developer B, 1 wash buffer, 1 concentrated wash solution.
- 96-well configuration: 1×96 enzyme-labeled plate, 2 seal films, 2 sealed bags, 1 standard, 1 standard diluent, 1 sample diluent, 1 developer A, 1 developer B, 1 wash buffer, 1 concentrated wash solution.
**Storage Instructions:**
All components should be stored at 2–8°C. Ensure all reagents are properly sealed after use to maintain stability.
**Sample Preparation and Handling:**
1. **Serum:** Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes. Collect the supernatant; if precipitate forms, re-centrifuge before testing.
2. **Plasma:** Use EDTA or sodium citrate as anticoagulant. Mix well for 10–20 minutes, then centrifuge for 20 minutes. Collect the supernatant carefully.
3. **Urine:** Collect in a sterile tube, centrifuge for 20 minutes, and collect the supernatant.
4. **Cell Culture Supernatant:** For secreted proteins, collect in a sterile tube and centrifuge. For intracellular components, lyse cells via repeated freeze-thaw cycles, then centrifuge again.
5. **Tissue Specimens:** Weigh the tissue, add PBS (pH 7.4), homogenize, centrifuge, and collect the supernatant. Store at 2–8°C after thawing.
6. **General Note:** Process samples as soon as possible after collection. If not tested immediately, store at -20°C, avoiding repeated freezing and thawing.
7. **Note:** Samples containing NaN3 should not be used, as it inhibits HRP activity.
This ELISA kit provides a reliable and efficient method for measuring mouse ALP levels, suitable for research applications in immunology, toxicology, and clinical diagnostics.
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