Human interleukin-15 (IL-15) ELISA test kit instruction manual

**Human Interleukin-15 (IL-15) ELISA Kit – Instructions for Use** This kit is intended for research purposes only and is designed to measure the concentration of human interleukin-15 (IL-15) in various biological samples such as serum, plasma, urine, cell culture supernatants, and other body fluids. The method employed is a double-antibody sandwich ELISA, which ensures high specificity and sensitivity for IL-15 detection. **Principle of the Assay** The assay utilizes a pre-coated microplate with a specific monoclonal antibody against IL-15. After incubation with the sample, the IL-15 present in the sample binds to the immobilized antibody. A horseradish peroxidase (HRP)-labeled secondary antibody is then added, forming a complex of antibody-antigen-enzyme-labeled antibody. Following washing steps, a TMB substrate is introduced, leading to a color change that is proportional to the amount of IL-15 in the sample. The reaction is stopped with an acidic solution, and the absorbance is measured at 450 nm using a microplate reader. The concentration of IL-15 in the sample is determined by comparing the OD values to a standard curve. **Kit Components** The kit includes: - Microtiter plate coated with anti-IL-15 antibody - Standard solutions (concentrated and diluted) - Enzyme-conjugated reagent - Substrate solutions (A and B) - Washing buffer (concentrated) - Sample diluent - Sealing film and storage bags **Storage Conditions** All components should be stored at 2–8°C. The shelf life is 6 months from the date of manufacture. **Sample Preparation** Proper handling of samples is crucial for accurate results. Serum and plasma should be obtained by centrifugation after clotting or anticoagulant treatment. Urine and cell culture supernatants should be centrifuged to remove debris. For tissue samples, homogenization in PBS followed by centrifugation is required. All samples must be processed promptly, and repeated freezing and thawing should be avoided. **Procedure Summary** 1. Prepare standard dilutions and load them into designated wells. 2. Add sample diluent and test samples to respective wells. 3. Incubate the plate at 37°C for 30 minutes. 4. Wash the plate thoroughly with the diluted washing buffer. 5. Add the HRP-labeled antibody and incubate again. 6. Add TMB substrate and develop the color for 15 minutes. 7. Stop the reaction with the stop solution. 8. Measure absorbance at 450 nm and calculate the IL-15 concentration. **Important Notes** - Allow all reagents to reach room temperature before use. - Avoid contamination by using separate pipettes for each step. - Always include blank wells and prepare a standard curve for accurate quantification. - Do not mix reagents from different batches. - Treat all waste materials as biohazardous. **Data Analysis** Plot the standard curve using the OD values versus known concentrations. Determine the sample concentration by interpolation or linear regression. Multiply the result by the dilution factor if applicable. **Performance Characteristics** - Sensitivity: 0.5 ng/L - Dynamic range: 0.5–10 ng/L - Intra- and inter-assay variability: <9% and <11%, respectively - Correlation coefficient (R²): ≥0.95 For best results, follow the instructions carefully and ensure proper calibration and quality control. This ELISA kit provides a reliable and efficient method for measuring IL-15 levels in human samples.

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