Duck Bcl-2 ELISA Kit

**Duck Bcl-2 ELISA Kit – For the Quantitative In Vitro Determination of Duck B-cell Leukemia/Lymphoma 2 in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids** *For Laboratory Research Use Only. Not Intended for Diagnostic or Therapeutic Procedures.* Before using this kit, please read the entire package insert carefully. This ELISA (Enzyme-Linked Immunosorbent Assay) is designed to quantitatively measure Duck Bcl-2 levels in various biological samples. The test works by detecting the presence of Bcl-2 through a colorimetric reaction, which is measured at 450 nm using a spectrophotometer. The kit includes a set of calibration standards to create a standard curve, allowing the operator to determine the concentration of Bcl-2 in unknown samples by comparing their optical density (OD) values to the standard curve. --- ### **Sample Collection and Storage** - **Serum**: Use a serum separator tube. Allow samples to clot for 2 hours at room temperature or overnight at 4°C. Centrifuge at 2000×g for 20 minutes. Remove the serum and assay immediately, or aliquot and store at -20°C. Avoid repeated freeze-thaw cycles. - **Plasma**: Collect using heparin as an anticoagulant. Centrifuge within 30 minutes of collection at 2000×g for 30 minutes at 2–8°C. Store at -20°C. Avoid freeze-thaw cycles. - **Cell Culture Supernatants, Tissue Homogenates, and Other Biological Fluids**: Centrifuge to remove particulates. Assay immediately or store at -20°C. Ensure no hemolysis or granules are present. --- ### **Materials Required (Not Supplied)** 1. Incubator set to 37°C 2. Microplate reader capable of measuring absorbance at 450 nm 3. Precision pipettes, disposable tips, and absorbent paper 4. Distilled or deionized water --- ### **Reagents Provided (Stored at 2–8°C)** | Reagent | 96 Determinations | 48 Determinations | |--------|-------------------|-------------------| | MicroELISA Strip Plate | 12×8 strips | 12×4 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 | **Note:** - Standard concentrations: 200, 100, 50, 25, 12.5, 6.25 ng/mL - If sample values exceed the highest standard, dilute with Sample Diluent. - Always use reagents provided by the manufacturer. - Do not thaw samples or reagents in a water bath. - Do not use beyond expiration date. - Use only deionized or distilled water. - Keep microtiter plate in its sealed bag until needed. - Discard used pipette tips properly. - Wear gloves and protective clothing during the procedure. - Treat all samples as potentially infectious. - Dispose of waste after 30 minutes of inactivation. - Substrate B contains 20% acetone—keep away from heat or flame. --- ### **Reagent Preparation and Storage** - **Wash Solution (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. Add 50 µL of standard or sample to each well (duplicate). Blank wells receive no addition. 2. Add 100 µL of HRP-conjugate reagent to all wells except blank. Cover with adhesive strip and incubate for 60 minutes at 37°C. 3. Wash the plate 4 times manually or using an automated washer. After final wash, invert and blot dry. 4. Add 50 µL of Chromogen A and 50 µL of Chromogen B to each well. Incubate for 15 minutes at 37°C, protecting from light. 5. Add 50 µL of Stop Solution to each well. The color should change from blue to yellow. Gently tap if uneven. 6. Read OD at 450 nm. Generate a standard curve by plotting average OD vs. concentration. Determine unknown concentrations by comparison. --- ### **Important Notes** - Each user should generate their own standard curve. - Intra-assay and inter-assay CV% < 15%. - Assay range: 6.25–200 ng/mL. - Sensitivity: <1.0 ng/mL. No cross-reactivity observed. - Storage: 2–8°C (frequent use); 6 months at -20°C. - Handle all reagents with care. Follow good laboratory practices. - Disposal: Inactivate viruses for 30 minutes before disposal. This kit is ideal for research purposes only and should not be used for clinical diagnosis. Always follow safety protocols when handling biological materials.

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