Human S-100B ELISA Kit

**Human S-100B ELISA Kit – For the Quantitative In Vitro Determination of Human Soluble Protein-100B Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Body Fluids** *For Laboratory Research Use Only. Not for Use in Diagnostic Procedures.* This kit is designed for the quantitative determination of human S-100B protein levels in various biological samples. The assay is based on the enzyme-linked immunosorbent assay (ELISA) principle, which involves a colorimetric reaction that is measured at 450 nm using a spectrophotometer. The intensity of the color produced is directly proportional to the concentration of S-100B in the sample. The kit includes a set of calibration standards, which are used to generate a standard curve by plotting optical density (OD) values against known concentrations of S-100B. This allows researchers to accurately determine the concentration of S-100B in unknown samples by comparing their OD readings to the standard curve. **Sample Collection and Storage:** - **Serum:** Collect using a serum separator tube. Allow clotting for 2 hours at room temperature or overnight at 4°C before centrifuging at 2000×g for 20 minutes. Store samples 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 samples are properly centrifuged and free from hemolysis or debris. **Materials Required but 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:** 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 Stripplate | 12x8 strips | 12x4 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 | **Important Notes:** - Standard concentrations: 640, 320, 160, 80, 40, 20 pg/mL. - If sample values exceed the highest standard, dilute with Sample Diluent. - Always use reagents provided by the manufacturer. - Allow all reagents and materials to reach room temperature (20–25°C) before use. - Do not thaw samples or reagents using water baths. - Do not use components beyond their expiration date. - Use only deionized or distilled water for dilutions. - Keep unused microtiter plate strips in their sealed pouch with desiccant at 2–8°C. - Use fresh pipette tips for each transfer to avoid contamination. - All blood-derived products should be treated as potentially infectious. - Follow proper disposal procedures for all waste materials. **Wash Solution Preparation (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 starting. It is recommended to 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 microtiter plate 4 times. Manual washing: aspirate contents, fill with wash solution, and repeat. Automated washing: follow manufacturer guidelines. 5. Add 50 µL of Chromogen Solution A and 50 µL of Chromogen Solution B to each well. Incubate for 15 minutes at 37°C, protected from light. 6. Add 50 µL of Stop Solution to each well. The color will change from blue to yellow. If uneven color appears, gently tap the plate. 7. Read OD at 450 nm. Generate a standard curve by plotting average OD values against standard concentrations. Determine sample concentrations by comparing their OD to the curve. **Performance Characteristics:** - Sensitivity: <20 pg/mL - Assay Range: 20–640 pg/mL - Intra-assay CV: <15% - Inter-assay CV: <15% - Cross-reactivity: No significant cross-reactivity observed **Storage:** - 2–8°C for frequent use; 6 months at -20°C. **Safety and Disposal:** - Liquid waste should be treated with sodium hypochlorite (final concentration 1.0%) and allowed to stand for 30 minutes before disposal. - Substrate B contains 20% acetone—keep away from heat and flame. - Handle all reagents carefully and follow good laboratory practices.

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