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 – Instructions for Use**
This ELISA kit is designed for the quantitative determination of human immunoreactive growth hormone (irGH) in biological samples such as serum, plasma, urine, cell culture supernatants, and tissue homogenates. The kit uses a sandwich ELISA method, which ensures high specificity and sensitivity for accurate detection.
**Kit Specifications:**
- 48-well or 96-well configuration
- Standard dilution: 1.5 mL × 1 bottle
- Enzyme standard reagent: 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- Storage conditions: 2–8°C
- Shelf life: 6 months from the date of receipt
**Kit Components:**
- Sealing film: 2 pieces (48-well) / 2 pieces (96-well)
- Standard: 0.5 mL × 1 bottle (2700 ng/L)
- Sample diluent: 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- Developer A: 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- Chromogen B: 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- Wash buffer (concentrated): 20 mL × 20 times (48-well) / 20 mL × 30 times (96-well)
- Stop solution: 3 mL × 1 bottle
- Anti-irGH coated microplate
**Principle of Operation:**
The ELISA kit utilizes a double-antibody sandwich technique. The microplate is pre-coated with anti-irGH antibodies. After adding the sample, irGH binds to the immobilized antibody. A horseradish peroxidase (HRP)-labeled secondary antibody is then added, forming an antibody-antigen-enzyme complex. TMB substrate is used for color development, and the reaction is stopped with a stop solution. The absorbance at 450 nm is measured, and the concentration of irGH is determined using a standard curve.
**Sample Preparation:**
- **Serum/Plasma:** Centrifuge at 2000–3000 rpm for 20 minutes after clotting or anticoagulant addition.
- **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant:** Centrifuge at 2000–3000 rpm for 20 minutes. For intracellular components, lyse cells by freezing/thawing before centrifugation.
- **Tissue:** Homogenize in PBS, centrifuge, and collect the supernatant.
**Storage & Handling:**
- Store all reagents at 2–8°C.
- Avoid repeated freeze-thaw cycles.
- Do not use samples containing NaN3, as it inhibits HRP activity.
**Procedure Summary:**
1. Prepare standard dilutions and load into designated wells.
2. Add sample diluent and test samples to respective wells.
3. Incubate at 37°C for 30 minutes.
4. Wash the plate 5 times with diluted washing buffer.
5. Add enzyme-labeled reagent 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 using a microplate reader.
9. Calculate concentrations using the standard curve.
**Notes:**
- Allow the kit to equilibrate at room temperature before use.
- Ensure proper pipetting accuracy and avoid cross-contamination.
- Always run standards in duplicate for reliable results.
- If sample OD exceeds the highest standard, perform a preliminary dilution.
**Performance:**
- Linear range: 0.2 IU/L – 6 IU/L
- Correlation coefficient (R²) ≥ 0.95
- Intra-batch CV < 9%, Inter-batch CV < 11%
**Service Commitment:**
We provide free technical support during working hours. Sample testing services are available upon request to help maximize experimental success.
**Important:** This kit is for research use only. Do not use for diagnostic purposes.