Human IGF-1 ELISA Kit

**Human IGF-1 ELISA Kit – For the Quantitative In Vitro Determination of Human Insulin-Like Growth Factor 1 (IGF-1) Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Body Fluids** *For Laboratory Research Use Only. Not for Use in Diagnostic Procedures.* This ELISA kit is designed for the quantitative measurement of human IGF-1 levels in various biological samples. The Stop Solution changes the color from blue to yellow, and the optical density is measured at 450 nm using a spectrophotometer. To determine the concentration of IGF-1 in the sample, the kit includes a set of calibration standards. These standards are run alongside the samples to generate a standard curve, allowing the operator to calculate the IGF-1 concentration based on the optical density readings. **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 centrifugation at 2000×g for 20 minutes. Store at -20°C. Avoid repeated freeze-thaw cycles. - **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids:** Centrifuge to remove particulates and assay immediately, or aliquot and store at -20°C. Ensure proper centrifugation and avoid hemolysis or granules in the sample. **Materials Required but Not Supplied:** 1. Incubator 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 (Stored at 2–8°C):** | Reagent | 96 Determinations | 48 Determinations | |--------|-------------------|-------------------| | MicroELISA Strip Plate | 12 x 8 strips | 12 x 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: 320, 160, 80, 40, 20, 10 ng/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. All components are calibrated for optimal performance. 2. Allow all reagents and materials 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. Use only deionized or distilled water. 5. Keep microtiter plates in sealed bags until needed. Store unused strips at 2–8°C with desiccant. 6. Use fresh pipette tips for each transfer to prevent cross-contamination. 7. Disposable gloves must be worn during the procedure. All blood-derived products should be treated as potentially infectious. 8. Dispose of all waste properly, ensuring at least 30 minutes of inactivation time for viruses. 9. Substrate solutions are prone to contamination. Handle with care. 10. Chromogen B contains 20% acetone; keep away from heat and open flames. 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 appropriate wells. Blank well receives no addition. 3. Add 100 µl of HRP-conjugate reagent to all wells except the blank. Cover with an adhesive strip and incubate for 60 minutes at 37°C. 4. Wash the plate 4 times manually or automatically. For manual washing, fill each well with 1X Wash Solution, aspirate, and repeat four times. After the final wash, invert and blot dry. 5. Add 50 µl of Chromogen A and 50 µl of Chromogen B to each well. Incubate for 15 minutes at 37°C, protecting from light. 6. Add 50 µl of Stop Solution to each well. The color will change from blue to yellow. If uneven, gently tap the plate. 7. Measure OD at 450 nm. Generate a standard curve by plotting average OD values against corresponding concentrations. **Data Interpretation:** 1. Calculate the mean OD for each standard and sample. Subtract the blank value from all readings. 2. Plot the standard curve with OD on the Y-axis and concentration on the X-axis. 3. Locate the OD value of the unknown sample on the Y-axis and draw a horizontal line to intersect the curve. Draw a vertical line down to the X-axis to find the concentration. 4. Variations in technique, timing, or kit age may affect results. Each user should generate their own standard curve. 5. Intra-assay and inter-assay CV% are less than 15%. 6. Assay range: 10 ng/ml to 320 ng/ml. 7. Sensitivity: <10 ng/ml. 8. Cross-reactivity: No significant cross-reactivity with other growth factors. 9. Storage: 2–8°C (for frequent use); 6 months at -20°C. **Important Notes:** - This kit is intended for research purposes only. - Follow all safety protocols when handling biological samples. - Always refer to the manufacturer’s instructions for full details. **Quote: "Accuracy and precision are key to reliable results."**

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