Chicken carbonic anhydrase (CA) elisa kit instruction manual

**Chicken Carbonic Anhydrase (CA) ELISA Kit – Instructions for Use** This ELISA kit is designed for the quantitative determination of Chicken Carbonic Anhydrase (CA) in biological samples such as serum, plasma, urine, cell culture supernatants, and tissue homogenates. The kit uses a sandwich ELISA method, where the target antigen binds to a coated antibody on the microplate, followed by detection with an HRP-conjugated secondary antibody. The enzymatic reaction produces a color change that correlates with the concentration of CA in the sample. **Kit Specifications:** - **Configuration:** 48-well or 96-well format - **Standard Dilution:** 1.5 mL × 1 vial - **Enzyme Standard Reagent:** 3 mL × 1 vial (48-well) / 6 mL × 1 vial (96-well) - **Storage Conditions:** 2–8°C - **Expiration Date:** 6 months from the date of receipt **Kit Components:** - Sealing Film: 2 pieces (48-well) / 2 pieces (96-well) - Standard: 2700 ng/L, 0.5 mL × 1 vial - Sample Diluent: 3 mL × 1 vial (48-well) / 6 mL × 1 vial (96-well) - Developer A: 3 mL × 1 vial (48-well) / 6 mL × 1 vial (96-well) - Chromogen B: 3 mL × 1 vial (48-well) / 6 mL × 1 vial (96-well) - Wash Buffer: 3 mL × 1 vial (48-well) / 6 mL × 1 vial (96-well) - Concentrated Wash Solution: 20 mL × 20 times (48-well) / 20 mL × 30 times (96-well) **Principle of Operation:** The kit employs a double-antibody sandwich ELISA technique. Purified anti-CA antibodies are immobilized on the microplate. After incubation with the sample, CA binds to the immobilized antibody. A horseradish peroxidase (HRP)-labeled secondary antibody is then added, forming a complex. TMB substrate is used for color development, which changes from blue to yellow upon acid termination. The absorbance at 450 nm is measured and correlated with the CA concentration using a standard curve. **Purpose:** To quantitatively measure the level of Carbonic Anhydrase (CA) in chicken serum, plasma, urine, and other biological fluids. **Sample Preparation:** - **Serum:** Allow blood to clot at room temperature for 10–20 minutes, then centrifuge at 2000–3000 rpm for 20 minutes. - **Plasma:** Use EDTA or sodium citrate as anticoagulant, mix, and centrifuge similarly. - **Urine:** Centrifuge at 2000–3000 rpm for 20 minutes. - **Cell Culture Supernatant:** Centrifuge after collection; for intracellular components, lyse cells by freezing and thawing. - **Tissue Homogenate:** Homogenize in PBS, centrifuge, and collect supernatant. **Notes:** - All samples should be processed as soon as possible after collection. - Avoid repeated freeze-thaw cycles. - Do not use samples containing NaN3, as it inhibits HRP activity. **Operation Steps:** 1. **Standard Dilution:** Prepare a serial dilution of the standard solution. 2. **Loading:** Add 40 μL of sample diluent and 10 μL of sample to each well. 3. **Incubation:** Seal the plate and incubate at 37°C for 30 minutes. 4. **Washing:** Rinse the plate 5 times with wash buffer. 5. **Enzyme Addition:** Add 50 μL of enzyme-labeled reagent to each well. 6. **Second Incubation:** Repeat incubation. 7. **Color Development:** Add 50 μL of TMB A and B, incubate at 37°C for 15 minutes. 8. **Stop Reaction:** Add 50 μL of stop solution. 9. **Measurement:** Read OD at 450 nm within 15 minutes. **Important Notes:** - Allow the kit to equilibrate at room temperature before use. - Ensure all reagents are properly mixed and handled. - Always prepare a standard curve and perform duplicate measurements. - Store unused reagents at 2–8°C and avoid cross-contamination. - Follow the manual strictly for accurate results. **Performance:** - Linear regression correlation coefficient (R) ≥ 0.95 - Intra-batch CV < 9%, Inter-batch CV < 11% - Detection range: 0.2 IU/L – 6 IU/L **Service Commitment:** - Free technical support during working hours. - Sample testing services available upon request. - Fast delivery after payment. **Storage and Handling:** - Store the kit at 2–8°C. - Do not use after the expiration date. This ELISA kit provides a reliable and sensitive method for measuring CA levels in chicken samples. Proper execution of the protocol ensures accurate and reproducible results.

3D-CAD Design

3D Shoulder Bags for Women,3D Crossbody Bags for Women,3D Women's Bags,3D-CAD Design Bags

NINGBO SUREWAY E-FASHION IMPORT AND EXPORT CO,.LTD , https://www.surewaybags.com