Human FETU-A ELISA Kit

**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 PROCEDURES.** Before starting the assay, please read this entire package insert carefully to ensure proper use and accurate results. --- ### **INTENDED USE AND TEST PRINCIPLE** This FETU-A ELISA Kit is intended for laboratory research use only and is not suitable for diagnostic or therapeutic purposes. The kit uses a sandwich ELISA format to detect and quantify FETU-A levels in biological samples. The test works by binding FETU-A in the sample to specific antibodies coated on the microtiter plate. After washing, a horseradish peroxidase (HRP)-conjugated secondary antibody is added, which binds to the captured FETU-A. A chromogenic substrate is then introduced, producing a color change that is proportional to the amount of FETU-A present. The reaction is stopped with a stop solution, and the absorbance is measured at 450 nm using a microplate reader. To determine the concentration of FETU-A in each sample, a standard curve is generated using known concentrations of FETU-A. The optical density (OD) values from the samples are compared to the standard curve to calculate their respective FETU-A levels. --- ### **SAMPLE COLLECTION AND STORAGE** - **Serum**: Use a serum separator tube. Allow the blood to clot for 2 hours at room temperature or overnight at 4°C before centrifuging at 2000×g for 20 minutes. Remove the serum and analyze immediately, or aliquot and store at -20°C. Avoid repeated freeze-thaw cycles. - **Plasma**: Collect using heparin as an anticoagulant. Centrifuge within 30 minutes of collection at 2000×g for 30 minutes at 2–8°C. Store at -20°C. Avoid repeated freeze-thaw cycles. - **Cell culture supernatants, tissue homogenates, and other biological fluids**: Centrifuge to remove particulates. Analyze immediately or aliquot and store at -20°C. Ensure no hemolysis or granules are present in the samples. --- ### **MATERIALS REQUIRED BUT NOT SUPPLIED** 1. Incubator set at 37°C 2. Microplate reader capable of measuring absorbance at 450 nm 3. Precision pipettes, disposable tips, and absorbent paper 4. Distilled or deionized water --- ### **REAGENTS PROVIDED** All reagents should be stored at 2–8°C. Check the expiration date on the label before use. | Reagent Name | 96 Determinations | 48 Determinations | |----------------------------------|-------------------|-------------------| | MicroELISA Strip Plate | 12×8 strips | 12×4 strips | | Standard (6 vials) | 0.5 ml/vial | 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 Solution A | 6.0 ml | 3.0 ml | | Chromogen Solution B | 6.0 ml | 3.0 ml | | Stop Solution | 6.0 ml | 3.0 ml | | Closure Plate Membrane | 2 | 2 | | User Manual | 1 | 1 | | Sealed Bags | 1 | 1 | *Note: Standard concentrations are 1000, 500, 250, 125, 62.5, 31.2 μg/mL.* --- ### **PRECAUTIONS AND GUIDELINES** 1. Only use reagents supplied by the manufacturer. 2. Allow all reagents and materials to reach room temperature (20–25°C) before use. Do not thaw using a water bath. 3. Do not use any reagent beyond its expiration date. 4. Use only deionized or distilled water for dilutions. 5. Keep unused microtiter plate strips in their sealed pouch with desiccant at 2–8°C. 6. Always use fresh disposable pipette tips to avoid cross-contamination. 7. All human-derived samples should be treated as potentially infectious. Follow good laboratory practices. 8. Dispose of all waste according to local regulations. Allow liquid to stand for 30 minutes before disposal to inactivate viruses. 9. Substrate solutions can be easily contaminated. Avoid exposure to heat or flame. 10. Handle all reagents with care and follow safety protocols. --- ### **ASSAY PROCEDURE** 1. Prepare all reagents and microtiter plate strips before beginning the assay. 2. Add 50 μL of standard or sample to the appropriate wells (excluding the blank well). Cover with adhesive strip and incubate for 60 minutes at 37°C. 3. Wash the microtiter plate 4 times using either manual or automated washing methods. 4. Add 50 μL of Chromogen Solution A and 50 μL of Chromogen Solution B to each well. Mix gently and incubate for 15 minutes at 37°C, protecting from light. 5. Add 50 μL of Stop Solution to each well. Read OD at 450 nm using a microplate reader. --- ### **DATA ANALYSIS** 1. Calculate the mean OD for each standard and sample. Subtract the blank OD value from all measurements. 2. Plot the average OD values against the corresponding FETU-A concentrations to generate a standard curve. 3. Determine the FETU-A concentration in each sample by locating its OD value on the Y-axis and drawing a line to intersect the standard curve. From the intersection point, draw a vertical line to the X-axis to find the corresponding concentration. --- ### **QUALITY CONTROL AND PERFORMANCE** - Intra-assay CV (%) and Inter-assay CV (%) are less than 15%. - Assay range: 31.2 μg/mL – 1000 μg/mL. - Sensitivity: Minimum detectable dose is typically less than 10 μg/mL. - Cross-reactivity: No significant cross-reactivity or interference observed. - Storage: Store at 2–8°C for frequent use; up to six months at -20°C. --- **For further assistance, contact the manufacturer or refer to the detailed user manual.**

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