Phosphorylated Adenosine Monophosphate Activated Protein Kinase （AMPK） BioAssay ELISA Kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of AMPK in human serum，plasma and other biological fluids.Detection Range：0.31-20ng/mlSensitivity：0.19ng/mlPrecision：Intra-Assay %CV： <10Inter-Assay %CV： <10Specificity： Recognizes human AMPK. No significant crossreactivity or interference between human AMPK and analogues was observed.Test Principle：The microtiter plate provided in this kit has been pre-coated with an antibody specific to AMPK. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to AMPK. Next，Avidin conjugated to Horseradish Peroxidase （HRP） is added to each microplate well and incubated. Free components are washed away. The substrate solution is added，only those wells that contain AMPK，biotin-conjugated antibody and enzyme-conjugated Avidin will apear blue in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid 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 AMPK. The concentration of AMPK in the sample is then determined by comparing the O.D. of the sample to the standard curve.Kit Components：*381892A： Microtiter Strips，1x96 wells，Pre-coated; ready to use*381892B： Standard，2x1vial 381892C： Standard Diluent，1x20ml*381892D： Biotin Detection，100X，1x120ul *381892E： HRP Conjugate，100X，1x120ul 381892F： Biotin Detection Diluent，1x14ml381892G： HRP Diluent，1x14ml381892H： Substrate，1x10ml381892K： Stop Solution，1x10ml381892L： Wash Buffer，25X，1x30ml Precaution：The Stop Solution （381892K） 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 *381892A，*381892B，*381892D and *381892E at -20°C. Store all the other components at 4°C. 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 37°C.3. Aspirate and add 100ul prepared Biotin Working Solution. Incubate 1 hour at 37°C.4. Aspirate and wash 3 times.5. Add 100ul prepared HRP Detection Working Solution. Incubate 30 minutes at 37°C.6. Aspirate and wash 5 times.7. Add 90ul Substrate. Incubate 15 minutes at 37°C.8. Add 50ul Stop Solution. Read at 450nm immediately.
