**Rat Growth Hormone Releasing Polypeptide (GHRP) ELISA Kit Instruction Manual**
**Kit Specifications:**
The Rat GHRP ELISA Kit is available in a 48-well or 96-well configuration. It includes the following components:
- **Standard Diluent:** 1.5 mL × 1 bottle
- **Enzyme Standard Reagent:** 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- **Sealing Film:** 2 pieces (48-well) / 2 pieces (96-well)
- **Standard:** 0.5 mL × 1 bottle, concentration of 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 Solution:** 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- **Stop Solution:** 3 mL × 1 bottle (48-well) / 6 mL × 1 bottle (96-well)
- **Concentrated Washing Solution:** 20 mL × 20 times × 1 bottle (48-well) / 20 mL × 30 times × 1 bottle (96-well)
**Storage Conditions and Expiration:**
- Store all components at 2–8°C.
- The kit has a shelf life of 6 months from the date of receipt.
**Purpose:**
This ELISA kit is designed for the quantitative determination of growth hormone-releasing polypeptide (GHRP) in rat serum, plasma, urine, cell culture supernatants, tissue homogenates, and other biological fluids.
**Principle of the Assay:**
The kit utilizes a sandwich ELISA method. A microplate is pre-coated with a specific antibody against GHRP. After adding the sample and standard, the target GHRP binds to the immobilized antibody. An HRP-conjugated secondary antibody is then added, forming a complex. After washing, TMB substrate is added, leading to a color change that is proportional to the GHRP concentration. The reaction is stopped, and absorbance is measured at 450 nm. The concentration of GHRP in the sample is determined by comparing its OD value to a standard curve.
**Sample Preparation and Handling:**
- **Serum/Plasma:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant:** Centrifuge after collection. For intracellular components, lyse cells using repeated freeze-thaw cycles.
- **Tissue Homogenate:** Homogenize in PBS, centrifuge, and collect the supernatant.
- **Storage:** Samples should be processed as soon as possible. If not used immediately, store at -20°C. Avoid repeated freezing and thawing.
**Notes and Recommendations:**
- Always equilibrate the kit to room temperature before use.
- Use a separate pipette for each step to avoid cross-contamination.
- Prepare a standard curve with duplicate wells. If the sample OD exceeds the highest standard, dilute the sample and re-test.
- Do not mix reagents from different batches.
- Keep the substrate away from light.
- All waste materials should be treated as biohazardous.
- Follow the manufacturer’s instructions strictly.
**Performance:**
- The kit demonstrates a high correlation coefficient (R ≥ 0.95) between the standard curve and expected concentrations.
- Intra-assay and inter-assay variability are <9% and <11%, respectively.
- Detection range: 0.2 IU/L – 6 IU/L.
**Service Commitment:**
- Free technical support during working hours.
- Sample testing services available upon request.
- Fast delivery after payment.
**Important:** This product is for research use only. Do not use it for diagnostic or therapeutic purposes.
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