The Coagulation Factor VIII (F8) BioAssay™ ELISA Kit (Human) is a sandwich enzyme immunoassay for the in vitro quantitative measurement of F8 in human plasma.
Detection Range:78-5,000pg/ml
Sensitivity:<30pg/ml
Precision:Intra-Assay: CV<10%Inter-Assay: CV<12%
Test Principle:The microtiter plate provided in this kit has been pre-coated with an antibody specific to F8. Standards or samples are added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to F8. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain F8, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of F8 in the sample is determined by comparing the O.D. of the sample to the standard curve.
Storage and Stability:Store *024233A, *024233B, *024233D and *024233E at -20°C. Store all the other components at 4°C. Unused kit is stable for 6 months. Once kit components are opened, it is highly recommended to use remaining reagents within 1 month provided this is within the expiration date of the kit. For maximum recovery of product, 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 2 hours at 37°C
3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37°C
4. Aspirate and wash 3 times
5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37°C
6. Aspirate and wash 5 times
7. Add 90ul TMB Substrate. Incubate 10-20 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately
