Human β-CGRP ELISA Kit

**Human β-CGRP ELISA Kit – For the quantitative in vitro determination of Human β-calcitonin gene-related peptide concentrations in serum, plasma, cerebrospinal fluid, tissue homogenate, and other body fluids. This kit is intended for laboratory research use only and not for diagnostic procedures.** The ELISA kit works by measuring the color intensity at 450 nm using a spectrophotometer. The test involves a series of steps where the sample's optical density (OD) is compared to a standard curve generated from known concentrations of β-CGRP. This allows for accurate quantification of the target protein in various biological matrices. **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 before centrifugation at 2000×g for 20 minutes. Remove 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 and assay immediately or store at -20°C. Ensure samples are properly centrifuged and free of hemolysis or debris. **Materials Required (Not Supplied):** 1. 37°C incubator 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 (Store at 2–8°C):** - MicroELISA Strip Plate (12×8 or 12×4 strips) - Standard (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 Solution A (6.0 ml or 3.0 ml) - Chromogen Solution B (6.0 ml or 3.0 ml) - Stop Solution (6.0 ml or 3.0 ml) - Closure Plate Membrane (2 units) - User Manual - Sealed Bags **Standard Concentrations:** 200, 100, 50, 25, 12.5, 6.25 pg/mL. If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay. **Precautions:** - Do not mix reagents from different kit lots. - Allow all reagents and samples to reach room temperature (20–25°C) before use. - Do not use beyond the expiration date. - Keep microtiter plates in their sealed bag until needed. Unused strips should be stored at 2–8°C with desiccant. - Wear gloves when handling biological samples due to potential infectious risk. - Dispose of waste after at least 30 minutes of inactivation. - Avoid contact with acid and sodium hypochlorite. - Chromogen Solutions A and B contain 20% acetone—keep away from heat or flame. **Reagent Preparation:** - Wash Solution (1X): Dilute 1 volume of 20X Wash Solution with 19 volumes of distilled or deionized water. Store at 2–8°C for 1 month. **Assay Procedure:** 1. Prepare all reagents before starting. Run standards and samples in duplicate. 2. Add 50 μL of standard or sample to each well. Blank well receives no addition. 3. Add 100 μL of HRP-conjugate reagent to all wells except blank. Cover and incubate for 60 minutes at 37°C. 4. Wash the plate 4 times manually or automatically. Ensure complete removal of liquid. 5. Add 50 μL of Chromogen A and 50 μL of Chromogen B. Incubate for 15 minutes at 37°C, protected from light. 6. Add 50 μL of Stop Solution. Gently mix if color change is uneven. 7. Measure OD at 450 nm. Plot average OD vs. concentration to generate the standard curve. **Interpretation:** - Calculate mean OD for each standard and sample. Subtract blank OD before interpretation. - Determine sample concentration by matching its OD to the standard curve. - Intra-assay and inter-assay CVs are <15%. - Assay range: 6.25–200 pg/mL. - Sensitivity: <1.0 pg/mL. - Cross-reactivity: No significant interference observed. - Storage: 2–8°C (frequent use); 6 months at -20°C. **Note:** Always follow good lab practices and handle all materials carefully. This kit is designed for research purposes only.

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