The MUC1 (Mucin 1) BioAssay™ ELISA Kit (Human) is a quantitative sandwich assay for the detection of MUC1 in human serum, plasma and other biological fluids.
Assay Principle:The microtiter plate provided in this kit has been pre-coated with an antibody specific to MUC1. Standards or samples are then added to the appropriate microtiter plate wells, followed by biotin-conjugated antibody specific to MUC1. 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 the sandwich complex consisting of MUC1, biotin-conjugated antibody and HRP-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 ± 2nm. The absorbance value is directly proportional to the concentration of MUC1. The concentration of MUC1 in the sample is determined by comparing the O.D. of the sample 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 Summary:1. Add 100ul standard or sample to each well and incubate for 90 minutes at 37°C 2. Aspirate the liquid
3. Add 100ul Antibody-Biotin working solution to each well and incubate for 60 minutes at 37°C4. Aspirate the liquid and wash plate 3 times
5. Add 100ul Avidin-HRP working solution to each well. Incubate for 30 minutes at 37°C6. Aspirate the liquid and wash plate 5 times
7. Add 90ul TMB substrate. Incubate for 15 minutes at 37°C8. Add 50ul Stop Solution. Read at 450nm immediately
