Human hemolytic complement (Hc) elisa kit instruction manual

**Human Hemolysis Complement (Hc) ELISA Kit Instruction Manual** **Kit Specifications:** - 48-well or 96-well configuration - Standard dilution: 1.5 ml × 1 bottle - Enzyme standard reagent: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96) - Storage conditions: 2–8°C for all components **Principle of the Assay:** This ELISA kit uses a sandwich immunoassay method to detect Human Hemolysis Complement (Hc) in serum, plasma, urine, and other biological fluids. The microplate is pre-coated with anti-Hc antibodies. After adding the sample, Hc binds to the immobilized antibody. A HRP-conjugated secondary antibody is then added, forming an antibody-antigen-enzyme complex. TMB substrate is used for color development, and the reaction is stopped with an acidic solution. The absorbance at 450 nm is measured, and the Hc concentration is calculated based on a standard curve. **Kit Composition:** - Sealing film: 2 pieces (48) / 2 pieces (96) - Standard: 2700 ng/L, 0.5 ml × 1 bottle - Enzyme standard: 1×48 / 1×96 - Sample diluent: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96) - Developer A: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96) - Chromogen B: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96) - Wash buffer: 3 ml × 1 bottle (48) / 6 ml × 1 bottle (96) - Concentrated wash solution: 20 ml × 20 times (48) / 20 ml × 30 times (96) **Storage & Shelf Life:** - Store at 2–8°C - Valid for 6 months from the date of manufacture **Sample Preparation Guidelines:** - **Serum:** Allow blood to clot at room temperature for 10–20 minutes, centrifuge at 2000–3000 rpm for 20 minutes. - **Plasma:** Use EDTA or sodium citrate as anticoagulant. Mix well and centrifuge as above. - **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes. - **Cell culture supernatant:** Centrifuge to remove debris. For intracellular components, lyse cells via freezing and thawing before centrifugation. - **Tissue:** Homogenize in PBS, centrifuge, and collect supernatant. **Precautions:** - Avoid repeated freeze-thaw cycles. - Do not use samples containing NaN3, as it inhibits HRP activity. - Always prepare a standard curve and perform duplicate measurements. - Keep all reagents away from light. - Follow instructions carefully for accurate results. **Operation Steps:** 1. Prepare standards by serial dilution. 2. Add sample diluent and test samples to the plate. 3. Incubate at 37°C for 30 minutes. 4. Wash the plate 5 times with diluted washing solution. 5. Add enzyme-labeled reagent and incubate again. 6. Add TMB substrate and incubate for 15 minutes. 7. Stop the reaction with stop solution. 8. Measure OD at 450 nm within 15 minutes. **Performance:** - Linear regression correlation coefficient (R²) ≥ 0.95 - Intra-assay CV < 9%, Inter-assay CV < 11% - Detection range: 0.2 IU/L – 6 IU/L **Purpose:** This kit is intended for research use only and is designed to quantify Human Hemolysis Complement (Hc) in various biological samples. **Service Commitment:** - Free technical support during working hours - Sample testing services available upon request - Fast delivery after payment **Note:** Always allow reagents to reach room temperature before use. If the enzyme reagent is opened, store it in a sealed bag. Avoid cross-contamination by using a new sealing film for each experiment. **Total Characters: 1100+**

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