Human MIG/CXCL9 ELISA Kit

**Human MIG/CXCL9 ELISA Kit – For the Quantitative In Vitro Determination of Human Monokine Induced by Interferon-Gamma Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids** **FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.** **INTENDED USE AND TEST PRINCIPLE** This Human MIG/CXCL9 ELISA Kit is specifically designed for laboratory research purposes and should not be used in diagnostic or therapeutic applications. The assay is based on a sandwich ELISA format, where the presence of MIG/CXCL9 in the sample is detected using specific antibodies. A stop solution is added to terminate the reaction, changing the color from blue to yellow. The optical density (OD) is measured at 450 nm using a spectrophotometer. Calibration standards are included in the kit to generate a standard curve, allowing for accurate quantification of MIG/CXCL9 levels in unknown samples. **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. 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 at 2000×g for 30 minutes at 2–8°C. Store at -20°C. Avoid repeated freezing and thawing. - **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids**: Centrifuge to remove particulates. Analyze immediately or store at -20°C. Ensure no hemolysis or contamination occurs during collection. **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. - MicroELISA Strip Plate: 12×8 strips (96 determinations), 12×4 strips (48 determinations) - Standards (6 vials): 0.5 ml/vial - Sample Diluent: 6.0 ml (96), 3.0 ml (48) - HRP-Conjugate Reagent: 10.0 ml (96), 5.0 ml (48) - 20X Wash Solution: 25 ml (96), 15 ml (48) - Chromogen Solution A: 6.0 ml (96), 3.0 ml (48) - Chromogen Solution B: 6.0 ml (96), 3.0 ml (48) - Stop Solution: 6.0 ml (96), 3.0 ml (48) - Closure Plate Membrane: 2 units - User Manual: 1 unit - Sealed Bags: 1 unit **NOTES** - Standard concentrations: 5000, 2500, 1250, 625, 312, 156 pg/ml - If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay. **PRECAUTIONS** 1. Do not mix reagents from different kit lots. Each component is calibrated for optimal performance. 2. Allow all reagents to reach room temperature (20–25°C) before use. Avoid using water baths for thawing. 3. Do not use reagents past their expiration date. 4. Only use deionized or distilled water for dilutions. 5. Keep microtiter plates in their sealed bags until use. Unused strips should be stored at 2–8°C with desiccant. 6. Use fresh disposable pipettes. Avoid contact with strong acids or sodium hypochlorite. 7. Handle all biological materials as potentially infectious. Wear gloves and follow good lab practices. 8. Dispose of all samples and waste properly. Add 1% sodium hypochlorite to liquid waste and let stand for 30 minutes before disposal. 9. Substrate solutions may be easily contaminated. Discard if they appear bluish. 10. Chromogen B contains 20% acetone—keep away from heat or flame. 11. Allow all reagents to reach room temperature before starting the assay. **REAGENT PREPARATION AND STORAGE** - **Wash Solution (1X)**: Dilute 1 volume of 20X Wash Solution with 19 volumes of deionized or distilled water. Store at 2–8°C for up to one month. **ASSAY PROCEDURE** 1. Prepare all reagents before beginning. Run standards and samples in duplicate. 2. Add 50 µl of standard or sample to each well. Blank well receives no addition. 3. Add 100 µl of HRP-conjugate reagent to all wells except the blank. Cover with adhesive strip and incubate for 60 minutes at 37°C. 4. Wash the plate 4 times manually or automatically. - Manual: Aspirate, fill with 1X Wash Solution, and repeat 4 times. - Automatic: Follow manufacturer instructions for washing. 5. 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. 6. Add 50 µl of Stop Solution. The color changes from blue to yellow. Gently tap the plate if discoloration is uneven. 7. Measure OD at 450 nm. Generate a standard curve by plotting average OD values against known concentrations. **RESULT INTERPRETATION** 1. Calculate the mean OD for each standard and sample. Subtract blank OD from all values. 2. Locate the OD value on the Y-axis and draw a horizontal line to intersect the standard curve. Read the corresponding concentration. 3. Variations in technique, time, or temperature can affect results. Each user should generate their own standard curve. 4. Intra-assay and inter-assay CV% are <15%. 5. Assay range: 156 pg/ml – 5000 pg/ml. 6. Sensitivity: <100 pg/ml. 7. Cross-reactivity: No significant cross-reactivity observed. 8. Storage: 2–8°C for frequent use; -20°C for long-term storage. **NOTES ON QUOTING** Please read all instructions carefully before performing the assay. Always follow safety protocols when handling biological samples.

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