This Anti-cyclic Citrullinated Peptide (CCP-Ab) BioAssay™ ELISA Kit is a sandwich ELISA for the in vitro quantitative determination of CCP-Ab concentrations in mouse serum, plasma and other biological fluids.
Specificity:Recognizes mouse CCP-Ab. No significant cross-reactivity or interference between mouse CCP-Ab and analogues was observed.
Test Principle:The micro ELISA plate provided in this kit has been pre-coated with an antigen specific to mouse CCP-Ab. Standards or samples are added to the micro ELISA plate wells and combined with the specific antigen. Then a biotinylated detection antigen specific for mouse CCP-Ab and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washedaway. The substrate solution is added to each well. Only those wells that contain mouse CCP-Ab, biotinylated detection antigen and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (O.D.) is measured spectrophotometrically at a wavelengthof 450nm ± 2nm. The O.D. value is proportional to the concentration of mouse CCP-Ab. You can calculate the concentration of mouse CCP-Ab in the samples by comparing the O.D. of the samples to the standard curve.
Storage and Stability:Unused kit is stable for 6 months after receipt. Store Microtiter Strips, Standard, and Antibody-Biotin at 4°C if kit will be used within 30 days; otherwise store at -20°C until expiration date. Store the rest of the components at 4°C. Protect HRP Conjugate and Substrate Reagent from light. For maximum recovery, centrifuge the original vials after thawing and prior to removing the cap.
Assay Procedure Summary:1. Prepare all reagents, samples and standards
2. Add 100ul standard or sample to each well. Incubate 90 minutes at 37°C
3. Aspirate and add 100ul prepared Biotin Working Solution. Incubate 1 hour at 37°C
4. Aspirate and wash 3 times
5. Add 100ul prepared HRP Detection Working Solution. Incubate 30 minutes at 37°C
6. Aspirate and wash 5 times
7. Add 90ul Substrate. Incubate 15 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
