**Monkey Alpha Interferon (IFN-α) ELISA Kit – Instructions for Use**
**Kit Specifications:**
This kit is available in 48-well or 96-well configurations. It includes:
- Standard dilution: 1.5 mL × 1 vial
- Enzyme standard reagent: 3 mL × 1 vial (for 48-well) / 6 mL × 1 vial (for 96-well)
- Sealing film: 2 pieces (for 48-well) / 2 pieces (for 96-well)
- Standard: 0.5 mL × 1 vial, concentration of 2700 ng/L
- Sample diluent: 3 mL × 1 vial (for 48-well) / 6 mL × 1 vial (for 96-well)
- Developer A: 3 mL × 1 vial (for 48-well) / 6 mL × 1 vial (for 96-well)
- Chromogen B: 3 mL × 1 vial (for 48-well) / 6 mL × 1 vial (for 96-well)
- Stop solution: 3 mL × 1 vial (for 48-well) / 6 mL × 1 vial (for 96-well)
- Concentrated washing solution: 20 mL × 20 times (for 48-well) / 20 mL × 30 times (for 96-well)
**Storage Conditions and Expiration:**
- Kit storage: 2–8°C
- Shelf life: 6 months from the date of manufacture
**Purpose:**
The Monkey Alpha Interferon (IFN-α) ELISA Kit is designed for the quantitative determination of IFN-α E in monkey serum, plasma, urine, cell culture supernatants, tissue homogenates, and other biological fluids.
**Principle of Operation:**
This kit employs a sandwich ELISA method. The microtiter plate is pre-coated with a specific antibody against IFN-α E. After adding the sample and enzyme-labeled antibody, a complex is formed. TMB substrate is added, and the reaction is stopped with an acidic solution. The color intensity is directly proportional to the IFN-α E concentration, which is measured at 450 nm using a microplate reader. A standard curve is plotted using known concentrations, and the unknown sample concentration is determined by comparison or linear regression.
**Sample Preparation Requirements:**
- **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, centrifuge similarly.
- **Urine:** Centrifuge for 20 minutes, collect supernatant.
- **Cell Culture Supernatant:** Centrifuge after collection; for intracellular components, lyse cells by repeated freeze-thaw and centrifuge.
- **Tissue:** Homogenize in PBS, centrifuge, and collect supernatant.
- **Storage:** Avoid repeated freezing and thawing. Store samples at -20°C if not tested immediately.
**Operation Steps:**
1. **Standard Dilution and Loading:** Prepare serial dilutions of the standard and load them into designated wells.
2. **Sample Loading:** Add diluted sample (final 5× dilution) to the test wells.
3. **Incubation:** Seal the plate and incubate at 37°C for 30 minutes.
4. **Washing:** Wash the plate 5 times with diluted washing buffer.
5. **Enzyme Labeling:** Add enzyme-conjugated antibody to all wells except blank.
6. **Second Incubation:** Repeat incubation at 37°C for 30 minutes.
7. **Color Development:** Add TMB reagents A and B, incubate for 15 minutes at 37°C.
8. **Stop Reaction:** Add stop solution to terminate the reaction.
9. **Measurement:** Read OD values at 450 nm within 15 minutes.
**Notes and Precautions:**
- Allow the kit to equilibrate at room temperature before use.
- Avoid cross-contamination by using a new sealing film for each experiment.
- Do not use samples containing NaN3, as it inhibits HRP activity.
- Ensure accurate pipetting and use a multichannel pipette for high-throughput testing.
- Always run standards in duplicate and prepare a standard curve for each assay.
- Follow all safety protocols when handling biological materials.
**Performance Characteristics:**
- Linearity: Correlation coefficient (R²) ≥ 0.95
- Intra-assay and inter-assay variability < 9% and < 11%, respectively
- Detection range: 0.2 IU/L – 6 IU/L
**Service Commitment:**
- Free technical support during working hours
- Free sample testing services to ensure optimal results
- Fast delivery upon payment
- English manual takes precedence in case of discrepancies
This kit is intended for research purposes only. Always follow the instructions carefully and validate results with a microplate reader.
Coffee Table,End Table,Round Side Table
Jiangmen Lihua Import & Export Trading Co., Ltd. Jiangmen Lihua Furniture., Ltd. , https://www.jmlihua.com