**Sheep IL-1 (IL-1) ELISA Kit – User Instructions**
This ELISA kit is designed for the quantitative determination of Interleukin-1 (IL-1) in various biological samples such as serum, plasma, urine, cell culture supernatants, and tissue homogenates. It utilizes a sandwich ELISA method, where the microplate is pre-coated with a specific antibody against sheep IL-1. The target antigen binds to the immobilized antibody, followed by the addition of HRP-conjugated detection antibody. A colorimetric reaction occurs upon substrate (TMB) addition, and the intensity of the color is directly proportional to the concentration of IL-1 in the sample.
**Kit Specifications:**
- **Configuration:** 48-well or 96-well format
- **Standard Dilution:** 1.5 mL × 1 vial
- **Enzyme Standard Reagent:** 3 mL × 1 vial (48-well) / 6 mL × 1 vial (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 vial (2700 ng/L)
- Enzyme Standard: 3 mL × 1 vial (48-well) / 6 mL × 1 vial (96-well)
- 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: 20 mL × 20 times or 20 mL × 30 times (diluted)
**Principle of Operation:**
The ELISA procedure involves binding of IL-1 to the coated antibody, followed by incubation with the HRP-labeled secondary antibody. After washing, TMB is added, and the reaction is stopped with an acidic solution. The absorbance at 450 nm is measured, and the IL-1 concentration is calculated based on a standard curve.
**Sample Preparation Guidelines:**
- **Serum/Plasma:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Cell Culture Supernatant:** Centrifuge at 2000–3000 rpm for 20 minutes.
- **Tissue Homogenate:** Homogenize in PBS, centrifuge, and collect supernatant.
**Precautions & Notes:**
- Avoid repeated freezing and thawing of samples.
- Do not use samples containing NaN₃, as it may inhibit HRP activity.
- Ensure all reagents are equilibrated to room temperature before use.
- Always run a standard curve and duplicate wells for accuracy.
- Use separate pipettes for each step to prevent cross-contamination.
- Store the plate in a sealed bag after opening to maintain performance.
**Performance Characteristics:**
- **Sensitivity:** 0.2 IU/L – 6 IU/L
- **Correlation Coefficient (R²):** ≥ 0.95
- **Intra-batch CV:** < 9%
- **Inter-batch CV:** < 11%
**Technical Support:**
For assistance with testing procedures, data interpretation, or sample analysis, please contact our technical support team during working hours. Free guidance and support are available to ensure optimal results.
**Storage & Handling:**
- Store the kit at 2–8°C.
- Avoid exposure to light and moisture.
- Do not mix reagents from different batches.
This ELISA kit is intended for research purposes only and should not be used for diagnostic or clinical applications.
Foshan Liqia Hardware Products Co., Ltd. , https://www.liqiamei.com