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Human IFN-α ELISA Kit
**Human IFN-α ELISA Kit – For the Quantitative In Vitro Determination of Human Interferon α in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Biological Fluids**
*For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.*
This kit is designed for the accurate and sensitive quantification of Human Interferon alpha (IFN-α) in various biological samples. The assay is based on a sandwich ELISA format, utilizing specific antibodies to capture and detect IFN-α. A standard curve is generated using a series of calibration standards, allowing for precise quantification of IFN-α levels in test samples.
The reaction is terminated by the addition of a Stop Solution, which changes the color from blue to yellow. The optical density (OD) is then measured at 450 nm using a microplate reader. By comparing the OD values of the samples to the standard curve, the concentration of IFN-α can be accurately determined.
**Sample Collection and Storage:**
- **Serum:** Use serum separator tubes. Allow samples to clot for 2 hours at room temperature or overnight at 4°C before centrifugation at 2000×g for 20 minutes. Remove the serum and assay immediately, or aliquot and store at -20°C. Avoid repeated freeze-thaw cycles.
- **Plasma:** Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g, 2–8°C, and store at -20°C. Avoid repeated freezing.
- **Cell Culture Supernatants, Tissue Homogenates, and Other Fluids:** Centrifuge to remove particulates. Assay immediately or store at -20°C. Ensure no hemolysis or granules are present.
**Materials Required but Not Supplied:**
- 37°C incubator
- Microplate reader capable of measuring absorbance at 450 nm
- Precision pipettes, disposable tips, and absorbent paper
- Distilled or deionized water
**Reagents Provided (Stored at 2–8°C):**
- 96-well strip plate (12 x 8 strips) / 48-well (12 x 4 strips)
- Standard (6 vials, 0.5 ml/vial)
- Sample Diluent (6.0 ml / 3.0 ml)
- HRP-Conjugate Reagent (10.0 ml / 5.0 ml)
- 20X Wash Solution (25 ml / 15 ml)
- Chromogen A (6.0 ml / 3.0 ml), Chromogen B (6.0 ml / 3.0 ml)
- Stop Solution (6.0 ml / 3.0 ml)
- Closure Plate Membrane (2 units)
- User Manual (1 copy)
- Sealed Bags (1 each)
**Precautions:**
- Do not mix reagents from different kits. Each kit is calibrated for optimal performance.
- Allow all reagents to reach room temperature (20–25°C) before use. Do not use water baths for thawing.
- Do not use reagents beyond their expiration date.
- Use only deionized or distilled water for dilution.
- Store unused strips in sealed bags with desiccant at 2–8°C.
- Use fresh pipette tips for each transfer to avoid contamination.
- Handle all samples with care, assuming potential infectious risk.
- Dispose of waste properly after inactivation for at least 30 minutes.
- Substrate solutions must be handled carefully; avoid exposure to heat or flame.
**Reagent Preparation:**
- Wash Solution (1X): Dilute 1 volume of 20X solution with 19 volumes of deionized water. Store at 2–8°C for 1 month.
**Assay Procedure:**
1. Prepare all reagents and add 50 µL of standard or sample to appropriate wells (excluding blank).
2. Cover with adhesive strip and incubate for 60 minutes at 37°C.
3. Wash the plate 4 times manually or using automated washers.
4. Add 50 µL of Chromogen A and B to each well, mix gently, and incubate for 15 minutes at 37°C.
5. Add 50 µL Stop Solution to each well and measure OD at 450 nm.
**Data Analysis:**
- Plot average OD values of standards vs. concentration.
- Subtract blank OD from all measurements.
- Calculate sample concentrations by interpolating from the standard curve.
- Intra-assay and inter-assay CV% < 15%.
**Assay Range:** 5 pg/mL – 160 pg/mL
**Sensitivity:** <1.0 pg/mL
**Cross-Reactivity:** No significant cross-reactivity observed.
**Storage:** 2–8°C (frequent use); 6 months at -20°C.
**Note:** If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay. Always read the entire package insert before use.