Foshan Liqia Hardware Products Co., Ltd. , https://www.liqiamei.com
Human immunoreactive growth hormone (irGH) elisa kit instruction manual
**Human Immunoreactive Growth Hormone (irGH) ELISA Kit – User Manual**
This ELISA kit is designed for the quantitative determination of human immunoreactive growth hormone (irGH) in various biological samples, including serum, plasma, urine, cell culture supernatants, and tissue homogenates. The kit is intended for research use only and must not be used for diagnostic or therapeutic purposes.
**Kit Specifications:**
- **Configuration:** 48-well or 96-well format
- **Standard Dilution:** 1.5 mL × 1 vial
- **Enzyme Standard Reagent:** 3 mL × 1 vial (for 48-well), 6 mL × 1 vial (for 96-well)
- **Storage Conditions:** 2–8°C
- **Shelf Life:** 6 months from the date of manufacture
**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, 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 Buffer: 3 mL × 1 vial (48-well), 6 mL × 1 vial (96-well)
- Concentrated Washing Solution: 20 mL × 20 times (48-well), 20 mL × 30 times (96-well)
**Principle of Operation:**
The kit uses a sandwich ELISA method. The microtiter plate is pre-coated with a specific antibody against irGH. After adding the sample, the target antigen binds to the immobilized antibody. An HRP-conjugated secondary antibody then binds to the complex, forming an antibody-antigen-enzyme conjugate. TMB substrate is added, and the reaction is stopped with a stop solution. The color intensity is measured at 450 nm, and the concentration is calculated based on a standard curve.
**Sample Preparation Guidelines:**
- **Serum:** Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes.
- **Plasma:** Use EDTA or sodium citrate as anticoagulant, mix well, and centrifuge similarly.
- **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Tissue Homogenate:** Weigh the tissue, add PBS, homogenize, and centrifuge.
**Precautions:**
- Avoid repeated freeze-thaw cycles.
- Do not use samples containing NaN3, as it may inhibit HRP activity.
- Always prepare a standard curve and run samples in duplicate.
- Store all components at 2–8°C and avoid exposure to light.
**Procedure Summary:**
1. Prepare standards by serial dilution.
2. Add samples and standards to the plate.
3. Incubate at 37°C for 30 minutes.
4. Wash the plate five times with diluted washing buffer.
5. Add enzyme-labeled reagent and incubate again.
6. Add TMB substrate and incubate for 15 minutes.
7. Stop the reaction with stop solution.
8. Measure OD at 450 nm using a microplate reader.
**Performance:**
- Correlation coefficient (R²) ≥ 0.95
- Intra-assay CV < 9%, Inter-assay CV < 11%
- Detection range: 0.2 IU/L – 6 IU/L
**Technical Support:**
Our team is available during working hours to assist with any questions or provide guidance. Free sample testing services are also available upon request to ensure optimal results.
**Storage & Expiry:**
- Store the kit at 2–8°C.
- Shelf life: 6 months from the date of receipt.
This manual is provided to ensure accurate and reliable results. Please follow the instructions carefully and refer to the original English version in case of discrepancies.