Human β2-GP1 ELISA Kit

**Human β2-GP1 ELISA Kit – For the Quantitative In Vitro Determination of Human β2 Glycoprotein 1 Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Biological Fluids** **FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.** This ELISA kit is designed for the quantitative measurement of Human β2-Glycoprotein 1 (β2-GP1) in various biological samples. The assay is based on a sandwich immunoassay principle using specific antibodies to capture and detect β2-GP1. 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 with a microplate reader. Calibration standards are included to generate a standard curve, which allows for accurate quantification of β2-GP1 levels in test samples. **Intended Use:** This kit is intended for laboratory research purposes only and is not suitable for diagnostic or therapeutic applications. **Test Principle:** The β2-GP1 ELISA Kit utilizes a competitive or sandwich immunoassay format. The sample or standard is incubated with immobilized antibodies, followed by detection with enzyme-conjugated secondary antibodies. After washing, a chromogenic substrate is added, producing a color change that correlates with the amount of β2-GP1 present. The reaction is stopped, and absorbance is measured at 450 nm. **Sample Collection and Storage:** - **Serum:** Collect using a serum separator tube. Allow clotting for 2 hours at room temperature or overnight at 4°C. Centrifuge at 2000×g for 20 minutes. Store at -20°C, avoiding repeated freeze-thaw cycles. - **Plasma:** Use heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g (2–8°C). Store at -20°C. - **Cell Culture Supernatants, Tissue Homogenates, and Other Body Fluids:** Centrifuge to remove particulates. Assay immediately or store at -20°C. Ensure no hemolysis or contamination occurs. **Materials Required but Not Supplied:** - Incubator at 37°C - Microplate reader capable of measuring absorbance at 450 nm - Precision pipettes, disposable tips, and absorbent paper - Distilled or deionized water **Reagents Provided (Stored at 2–8°C):** - Microtiter Strip Plate (12×8 or 12×4 strips) - Standards (6 vials, 0.5 ml/vial) - Sample Diluent (6.0 ml or 3.0 ml) - HRP-Conjugate Reagent (10.0 ml or 5.0 ml) - 20X Wash Solution (25 ml or 15 ml) - Chromogen Solutions A & B (6.0 ml each) - Stop Solution (6.0 ml each) - Closure Plate Membrane (2 units) - User Manual and Sealed Bags (1 each) **Important Notes:** - Standard concentrations: 16, 8, 4, 2, 1, 0.5 ng/ml. - If sample values exceed the highest standard, dilute with sample diluent. - Only use reagents provided by the manufacturer. - Allow all reagents to reach room temperature (20–25°C) before use. - Do not thaw samples or reagents using a water bath. - Avoid using expired components. - Use only deionized or distilled water for dilutions. - Keep unused strip wells in sealed bags with desiccant. - Use fresh pipette tips for each transfer to prevent cross-contamination. **Washing Procedure:** - Manual Washing: Aspirate plate contents, fill with 1X wash solution, aspirate again. Repeat four times. Dry by blotting. - Automated Washing: Use 1X Wash Buffer, aspirate and wash four times. Adjust brush settings for optimal performance. **Assay Procedure:** 1. Prepare all reagents before starting. 2. Add 50 µl of standard or sample to appropriate wells. Cover with adhesive strip and incubate at 37°C for 60 minutes. 3. Wash plate 4 times. 4. Add 50 µl of Chromogen A and B, incubate for 15 minutes at 37°C. 5. Add 50 µl Stop Solution. Measure OD at 450 nm. 6. Plot standard curve using average OD values. Calculate sample concentrations by comparing to the curve. **Performance Characteristics:** - Intra-assay CV <15%, Inter-assay CV <15%. - Assay range: 0.5 ng/mL – 16 ng/mL. - Sensitivity: <0.1 ng/mL. - Cross-reactivity: No significant cross-reaction observed. **Storage:** - Store at 2–8°C for frequent use; for long-term storage, keep at -20°C. - Shelf life: 6 months at -20°C. **Safety and Disposal:** - All samples should be treated as potentially infectious. - Dispose of liquids after at least 30 minutes of inactivation. - Handle all reagents carefully, especially those containing acetone. - Follow standard lab safety protocols. **Note:** This document must be read in full before performing the assay. Always refer to the user manual for detailed instructions and troubleshooting.

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