Chicken ACTH ELISA Kit

**Chicken ACTH ELISA Kit – For the quantitative in vitro determination of Chicken adrenocorticotropic hormone (ACTH) concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other body fluids. Intended for laboratory research use only. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES.** Before using this product, please read this entire package insert carefully. This kit is designed to measure ACTH levels through an Enzyme-Linked Immunosorbent Assay (ELISA). The principle of the assay involves a competitive binding reaction between the ACTH in the sample and a labeled ACTH analog, with the optical density (OD) measured at 450 nm. A standard curve is generated using known ACTH concentrations, which allows for accurate quantification of unknown samples. **Storage Instructions:** Store all samples at -20°C. Avoid repeated freeze-thaw cycles. For cell culture supernatants, tissue homogenates, and other biological fluids, centrifuge to remove particulates and assay immediately or aliquot and store at -20°C. Ensure that samples are properly centrifuged and free from hemolysis or debris. **Materials Required but Not Supplied:** 1. Incubator set at 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 (All stored at 2–8°C):** - 96-well microplate (12×8 strips) - Standard (6 vials, 0.5 mL/vial) – Concentrations: 80, 40, 20, 10, 5, 2.5 pg/mL - 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 units) - User Manual (1 copy) - Sealed Bags (1 unit) **Precautions:** - Do not mix reagents from different kit lots. - Allow all reagents to reach room temperature (20–25°C) before use. - Do not use reagents past their expiration date. - Only use deionized or distilled water for dilution. - Keep unused microplate strips in sealed bags with desiccant. - Use fresh pipette tips for each transfer to prevent contamination. - Handle all biological samples as potentially infectious. - Dispose of all waste according to local regulations. Liquid waste should be treated with sodium hypochlorite (final concentration 1.0%) and left for at least 30 minutes before disposal. **Reagent Preparation:** Dilute 1 volume of 20X Wash Solution with 19 volumes of deionized or distilled water. Store the 1X solution at 2–8°C for up to one month. **Assay Procedure:** 1. Prepare all reagents before starting. 2. Add 50 µL of standard or sample to appropriate wells (excluding blank well), cover with adhesive strip, and incubate for 60 minutes at 37°C. 3. Wash the plate 4 times manually or automatically. 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. If uneven, gently tap the plate. 6. Read OD at 450 nm within 15 minutes using a microplate reader. **Calculation of Results:** 1. Plot average OD values of standards vs. concentrations on graph paper or software. 2. Subtract blank OD from all readings. 3. Locate sample OD on Y-axis, draw horizontal line to the curve, then vertical line to X-axis to determine ACTH concentration. 4. Intra- and inter-assay CVs are <15%. 5. Assay range: 2.5–80 pg/mL. 6. Sensitivity: <1.0 pg/mL. 7. Cross-reactivity: No significant interference observed. **Storage:** - 2–8°C for frequent use; - -20°C for long-term storage (up to 6 months). **Important Note:** This kit is intended for research purposes only. It is not approved for diagnostic use. Always follow proper safety protocols when handling biological materials.

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