The Mouse Occludin (OCLN) ELISA Kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of OCLN in mouse tissue homogenates and other biological fluids.
Detection Range:0.312-20ng/ml
Sensitivity:0.117ng/ml
Test Principle:The microtiter plate provided in this kit has been pre-coated with an antibody specific to OCLN. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody preparation specific for OCLN. 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 OCLN, 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 OCLN in the sample is then determined by comparing the O.D. of the sample to the standard curve.
Kit Components:*027216A: Microtiter Plate, 1x96wells, Pre-coated, ready to use*027216B: Standard, 2x1vial 027216C: Standard Diluent, 1x20ml*027216D: Detection Reagent A (green), 1x120ul *027216E: Detection Reagent B (red), 1x120ul 027216F: Assay Diluent A, 2x, 1x6ml027216G: Assay Diluent B, 2x, 1x6ml027216H: TMB Substrate, 1x9ml027216K: Stop Solution, 1x6ml027216L: Wash Buffer, 30X, 1x20ml
Storage and Stability:Store *027216A, *027216B, *027216D and *027216E 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. 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 Substrate Solution. Incubate 15-25 minutes at 37°C
8. Add 50ul Stop Solution. Read at 450nm immediately.
