Pet Ear Wipes,Cleaning Wipe For Dog Ears,Ear Cleaning Powder For Dogs Cats Ear,Ear Cleaning For Dogs ShangHai Youhang Technology Co.,LTD , https://www.yhecoclean.com
Fish elisa kit, adrenomedullin (ADM) enzyme-free technology
**Fish Adrenomedullin (ADM) ELISA Kit – Research Use Only**
This Fish Adrenomedullin (ADM) Enzyme-Linked Immunosorbent Assay (ELISA) Kit is designed for the quantitative determination of ADM in fish serum, plasma, and other biological fluids. It utilizes enzyme-free technology to ensure high sensitivity and accuracy. This kit is ideal for researchers working in aquatic biology, immunology, and environmental health studies.
**Principle of the Assay**
The kit employs a double-antibody sandwich method. A microtiter plate is pre-coated with purified fish ADM antibodies. After adding the sample, ADM binds to the immobilized antibody. An HRP-labeled secondary antibody then recognizes the captured ADM, forming an antibody-antigen-enzyme complex. Following washing steps, TMB substrate is added, and the reaction is stopped with sulfuric acid. The color intensity, measured at 450 nm, correlates directly with ADM concentration in the sample.
**Kit Components**
- 1×48 or 1×96 wells (depending on configuration)
- Standard: 18 ng/L (0.5 mL × 1 bottle)
- Standard Diluent: 1.5 mL × 1 bottle
- HRP-Conjugate Reagent: 3 mL × 1 bottle
- Sample Diluent: 3 mL × 1 bottle
- Chromogen A & B: 3 mL × 1 bottle each
- Stop Solution: 3 mL × 1 bottle
- Wash Solution (20×): 20 mL × 1 bottle (for 20–30 dilutions)
- Sealing Film & Plate Strip: As per kit configuration
**Storage Conditions**
All components should be stored at 2–8°C. The kit has a shelf life of 6 months from the date of manufacture.
**Sample Preparation Guidelines**
- **Serum**: Allow blood to clot at room temperature for 10–20 minutes, centrifuge at 2000–3000 rpm for 20 minutes.
- **Plasma**: Use EDTA or sodium citrate as anticoagulant. Mix for 10–20 minutes, centrifuge similarly.
- **Urine**: Collect in sterile tubes, centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant**: Centrifuge at 2000–3000 rpm for 20 minutes. For intracellular components, lyse cells via freeze-thaw cycles before centrifugation.
- **Tissue Samples**: Homogenize in PBS (pH 7.4), centrifuge, and collect supernatant.
**Assay Procedure**
1. Prepare standard dilutions in a serial manner (12 ng/L to 1 ng/L).
2. Add 40 μL sample diluent and 10 μL sample to each well (final 5-fold dilution).
3. Incubate at 37°C for 30 minutes.
4. Wash 5 times with diluted wash buffer.
5. Add 50 μL HRP-conjugated reagent to each well.
6. Incubate again at 37°C for 30 minutes.
7. Add 50 μL each of Chromogen A and B, incubate for 15 minutes at 37°C.
8. Stop the reaction with 50 μL stop solution.
9. Measure OD at 450 nm within 15 minutes.
**Important Notes**
- Avoid repeated freeze-thaw cycles.
- Do not use samples containing NaN3 due to HRP inhibition.
- Always prepare a standard curve and run duplicates for accurate results.
- Store unused plates in sealed bags after opening.
- Keep all reagents away from light during preparation and testing.
**Performance**
- Sensitivity: 0.5 ng/L – 16 ng/L
- Correlation coefficient (R²) ≥ 0.990
- Intra-batch CV < 9%, Inter-batch CV < 11%
This kit provides a reliable and efficient method for ADM detection in aquatic species, supporting research in physiology, disease biomarkers, and environmental monitoring. Always follow the manufacturer’s instructions for optimal performance.