Canine ISR-β ELISA Kit

**Canine ISR-β ELISA Kit – For Quantitative In Vitro Determination of Canine Insulin Receptor β in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Body Fluids** *For Laboratory Research Use Only. Not Intended for Diagnostic or Therapeutic Purposes.* This kit is designed for the quantitative determination of Canine Insulin Receptor Beta (ISR-β) in various biological samples using a sandwich ELISA method. The assay relies on specific antibodies that bind to ISR-β, followed by a colorimetric detection system. The reaction is terminated with a stop solution, and the optical density (OD) is measured at 450 nm. A standard curve is generated using known concentrations of ISR-β, allowing for accurate quantification of unknown samples. **Intended Use** The Canine ISR-β ELISA Kit is intended for laboratory research use only and is not suitable for diagnostic or clinical procedures. It provides a reliable and sensitive method for measuring ISR-β levels in biological fluids. **Principle of the Assay** The ELISA procedure involves incubating samples and standards in microtiter wells coated with anti-ISR-β antibodies. After washing, a horseradish peroxidase (HRP)-conjugated secondary antibody is added, which binds to the captured ISR-β. A chromogenic substrate is then used to produce a color change, which is proportional to the amount of ISR-β present. The reaction is stopped, and the OD is read at 450 nm. The concentration of ISR-β in the sample is determined by comparing its OD value 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 centrifugation at 2000×g for 20 minutes. Store at -20°C. Avoid repeated freeze-thaw cycles. - **Plasma**: Collect in anticoagulant tubes and process within 2 hours. Store at -20°C. - **Cerebrospinal Fluid (CSF)**: Centrifuge immediately after collection and store at -20°C. - **Tissue Homogenate**: Prepare fresh or store at -20°C. Ensure no hemolysis or particulate matter is present. - **Cell Culture Supernatant**: Centrifuge to remove debris and use immediately or store at -20°C. **Materials Required (Not Included)** - Incubator set 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 (Store at 2–8°C)** - Microplate (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) - Closure Plate Membrane (2 pieces) - User Manual and Sealed Bags **Precautions** - Do not mix reagents from different kits. - Allow all reagents to reach room temperature before use. - Do not use beyond expiration date. - Use only deionized or distilled water for dilutions. - Avoid contamination by using fresh pipette tips. - Handle all biological materials as potentially infectious. - Dispose of waste using 1% sodium hypochlorite for at least 30 minutes before disposal. - Keep substrate solutions away from heat and flame. **Assay Procedure** 1. Prepare all reagents and add 50 µL of standards or samples to appropriate wells. 2. Incubate for 60 minutes at 37°C. 3. Wash the plate 4 times with 1X Wash Solution. 4. Add 50 µL of Chromogen A and B to each well, incubate for 15 minutes at 37°C. 5. Add 50 µL of Stop Solution and measure OD at 450 nm. 6. Plot the standard curve and calculate sample concentrations. **Performance Characteristics** - Sensitivity: <1.0 U/ml - Range: 7.5–240 U/ml - Intra-assay CV: <15% - Inter-assay CV: <15% - Cross-reactivity: No significant interference observed **Storage** - 2–8°C for frequent use; up to 6 months at -20°C. Always read the full instructions before performing the assay.

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