**Mouse Parathyroid Hormone (PTH) ELISA Kit – Instructions for Use**
This ELISA kit is designed for the quantitative determination of mouse parathyroid hormone (PTH) in biological samples such as serum, plasma, urine, cell culture supernatants, and tissue homogenates. The kit is intended for research use only.
**Kit Specifications:**
- 48-well or 96-well configuration
- Enzyme standard reagent: 1.5 mL × 1 vial (for 48-well), 3 mL × 1 vial (for 96-well)
- Standard dilution: 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- Storage: 2–8°C
**Kit Components:**
- Sealing film: 2 pieces (48-well), 2 pieces (96-well)
- Standard: 0.5 mL × 1 vial, 2700 ng/L
- Enzyme standard: 1×48 or 1×96
- Sample diluent: 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- Developer A: 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- Chromogen B: 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- Wash solution: 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- Concentrated wash solution: 20 mL × 20 times or 20 mL × 30 times (per bottle)
**Principle of Operation:**
The kit uses a double-antibody sandwich ELISA method. Purified anti-mouse PTH antibody is coated on microtiter plate wells. After incubation with the sample, HRP-labeled PTH antibody binds to the captured antigen, forming an immune complex. TMB substrate is added, and the color develops under the action of HRP. The reaction is stopped with a stop solution, and the absorbance at 450 nm is measured. The concentration of PTH in the sample is calculated using a standard curve.
**Purpose:**
To quantify PTH levels in various biological specimens for research purposes.
**Storage Conditions:**
- Kit storage: 2–8°C
- Shelf life: 6 months from the date of manufacture
**Sample Preparation:**
1. **Serum:** Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes.
2. **Plasma:** Use EDTA or sodium citrate as anticoagulant. Mix and centrifuge similarly.
3. **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
4. **Cell Culture Supernatant:** Centrifuge after collection; for intracellular components, lyse cells by freeze-thaw cycles.
5. **Tissue Homogenate:** Weigh the tissue, add PBS, homogenize, and centrifuge.
6. **Storage:** Samples should be processed as soon as possible. If not tested immediately, store at -20°C. Avoid repeated freezing and thawing.
**Important Notes:**
- Do not use samples containing NaN3, as it may inhibit HRP activity.
- Always prepare a standard curve and run duplicates for accuracy.
- Ensure all reagents are at room temperature before use.
- Discard any unused reagents after opening.
- Handle all waste materials as biohazardous.
**Procedure Summary:**
1. Dilute standards and load into designated wells.
2. Add sample diluent and test samples.
3. Incubate at 37°C for 30 minutes.
4. Wash the plate 5 times with diluted wash solution.
5. Add enzyme conjugate and incubate again.
6. Add TMB substrate and develop color for 15 minutes.
7. Stop the reaction with stop solution.
8. Measure OD at 450 nm within 15 minutes.
**Performance:**
- Correlation coefficient (R) ≥ 0.95
- Intra-assay CV < 9%, Inter-assay CV < 11%
- Detection range: 0.2 IU/L – 6 IU/L
**Technical Support:**
Free technical assistance is available during working hours. For optimal results, we recommend testing your samples with our expert team.
**Safety and Compliance:**
All samples, reagents, and waste should be handled as infectious materials. Follow proper biosafety protocols.
**Warranty and Service:**
We guarantee the quality of our products. If you have any questions, please contact us for support. Our goal is to ensure your experiments yield accurate and reliable results.
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