The Human N-Acetylgalactosaminidase (NAGa) ELISA Kit is a sandwich enzyme immunoassay for the in vitro quantitative measurement of NAGa in human serum, plasma, tissue homogenates and other biological fluids. Detection Range: 0.312-20ng/ml Sensitivity: 0.128ng/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 NAGa. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to NAGa. 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 NAGa, 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 NAGa in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *026993A: Microtiter Plate, 1x96 wells, Pre-coated, ready to use *026993B: Standard, 2x1vial 026993C: Standard Diluent, 1x20ml *026993D: Detection Reagent A (green), 1x120ul *026993E: Detection Reagent B (red), 1x120ul 026993F: Assay Diluent A, 1x12ml 026993G: Assay Diluent B, 1x12ml 026993H: TMB Substrate, 1x9ml 026993K: Stop Solution, 1x6ml 026993L: Wash Buffer, 30X, 1x20ml Precaution: The Stop Solution (026993K) suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. Storage and Stability: Store *026993A, *026993B, *026993D and *026993E at -20C. Store all the other components at 4C. 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 37C. 3. Aspirate and add 100ul prepared Detection Reagent A. Incubate 1 hour at 37C. 4. Aspirate and wash 3 times. 5. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37C. 6. Aspirate and wash 5 times. 7. Add 90ul TMB Substrate. Incubate 15-25 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately.
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