The Mucin 2 (MUC2) BioAssay(TM) ELISA Kit is a sandwich ELISA for in vitro quantitative measurement of MUC2 in human serum, plasma and other biological fluids. Detection Detection Range: 0.31-20ng/ml Sensitivity: 0.19ng/ml Precision: Intra-Assay %CV: <10% Inter-Assay %CV: <10% Specificity: Recognizes human MUC2. No significant crossreactivity or interference between human MUC2. and analogues was observed. Test Principle: The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to human MUC2. Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for human MUC2 and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain human MUC2, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450nm 2nm. The OD value is proportional to the concentration of human MUC2. The concentration of human MUC2 in samples can be calculated by comparing the OD of the samples to the standard curve. Kit Components: *383761A: Microtiter Strips, 1x96 wells (8x12wells), Pre-coated, ready to use *383761B: Standard, 2x1vial 383761C: Standard Diluent, 1x20ml *383761D: Biotin Detection, 100X, 1x120ul *383761E: HRP Conjugate, 100X, 1x120ul 383761F: Biotin Detection Diluent, 1x14ml 383761G: HRP Diluent, 1x14ml 383761H: Substrate, 1x10ml 383761K: Stop Solution, 1x10ml 383761L: Wash Buffer, 25X, 1x30ml Precaution: The Stop Solution (383761K) included for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. Storage and Stability: Store *383761A, *383761B, *383761D and *383761E at -20C. Store all the other components at 4C. Unused kit is stable for 6 months after receipt. 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 90 minutes at 37C. 3. Aspirate and add 100ul prepared Biotin Working Solution. Incubate 1 hour at 37C. 4. Aspirate and wash 3 times. 5. Add 100ul prepared HRP Detection Working Solution. Incubate 30 minutes at 37C. 6. Aspirate and wash 5 times. 7. Add 90ul Substrate. Incubate 15 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately.
