Human FETU-A ELISA Kit
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**Human FETU-A ELISA Kit – For the Quantitative In Vitro Determination of Human FETU-A Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids**
**FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES.**
This ELISA kit is specifically designed for the quantitative measurement of Human FETU-A levels in various biological samples. The assay is based on the enzyme-linked immunosorbent assay (ELISA) principle, where a colorimetric reaction occurs in proportion to the concentration of FETU-A present. The reaction is terminated with a stop solution, and the optical density (OD) is measured at 450 nm using a microplate reader.
To ensure accurate results, the kit includes a set of standard solutions that are used to generate a standard curve. This allows for precise quantification of FETU-A in unknown samples by comparing their OD values against the standard curve.
**Sample Collection and Storage:**
- **Serum:** Use a serum separator tube. Allow samples to clot for 2 hours at room temperature or overnight at 4°C before centrifuging at 2000×g for 20 minutes. Store aliquots at -20°C and avoid repeated freeze-thaw cycles.
- **Plasma:** Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g and store at -20°C.
- **Tissue Homogenate and Other Fluids:** Centrifuge to remove particulates, then store at -20°C. Avoid hemolysis or contamination.
**Materials Required (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):**
- Microtiter strip plate (12×8 or 12×4 strips)
- Standard (6 vials, 0.5 mL/vial)
- Sample diluent (6.0 mL or 3.0 mL)
- HRP-conjugated reagent (10.0 mL or 5.0 mL)
- 20X Wash solution (25 mL or 15 mL)
- Chromogen A & B (6.0 mL or 3.0 mL each)
- Stop solution (6.0 mL or 3.0 mL)
- Closure plate membrane (2 units)
- User manual and sealed bags
**Important Notes:**
- Standard concentrations: 1000, 500, 250, 125, 62.5, 31.2 μg/mL.
- If sample values exceed the highest standard, dilute with sample diluent and repeat the test.
- Only use reagents provided by the manufacturer.
- All reagents should be brought to room temperature (20–25°C) before use.
- Do not thaw samples or reagents in a water bath.
- Avoid using expired reagents.
- Use only deionized or distilled water for dilutions.
- Keep unused strips in the sealed bag with desiccant.
- Use fresh pipette tips to prevent cross-contamination.
- Treat all blood-derived materials as potentially infectious.
- Follow proper disposal procedures, including allowing liquid to stand for 30 minutes before disposal to inactivate viruses.
- Avoid exposure of chromogen solutions to heat or flame.
**Assay Procedure:**
1. Prepare all reagents before starting the assay.
2. Add 50 µL of standard or sample to the appropriate wells, cover with an adhesive strip, and incubate for 60 minutes at 37°C.
3. Wash the plate 4 times using either manual or automated methods.
4. Add 50 µL of Chromogen A and 50 µL of Chromogen B to each well, mix gently, and incubate for 15 minutes at 37°C, protected from light.
5. Add 50 µL of Stop Solution to each well and measure OD at 450 nm.
**Data Interpretation:**
- Plot the average OD values of the standards on a graph (Y-axis vs. X-axis).
- Determine the FETU-A concentration in samples by comparing their OD values to the standard curve.
- Ensure that each user generates their own standard curve due to potential variations in technique or environmental conditions.
**Performance Characteristics:**
- Intra-assay CV (%) and Inter-assay CV (%) < 15%
- Assay range: 31.2–1000 μg/mL
- Sensitivity: <10 μg/mL
- Cross-reactivity: No significant cross-reactivity with other proteins
- Storage: 2–8°C (for frequent use), 6 months at -20°C
**Safety and Handling:**
- Always follow good laboratory practices.
- Dispose of all waste materials properly.
- Handle all reagents with care and wear appropriate personal protective equipment (PPE).
**Note:** This product is intended for research purposes only and should not be used in diagnostic or clinical settings. Always read the entire package insert before use.