Peroxisome Proliferator Activated Receptor Gamma Coactivator 1 Alpha (PPARgC1a) BioAssay(TM) ELISA Kit (Mouse) is a sandwich enzyme immunoassay for in vitro quantitative measurement of PPARgC1a in mouse tissue homogenates, cell lysates and other biological fluids. Detection Range: 0.156-10ng/ml Sensitivity: 0.057ng/ml Intra-Assay: CV: <10% Inter-Assay: CV: <12% Kit Components: *027388A: Microtiter Strips, 1x96 wells, Pre-coated, ready to use *027388B: Standard, 2x1vial 027388C: Standard Diluent, 1x20ml *027388D: Detection Reagent A, 1x120ul *027388E: Detection Reagent B, 1x120ul 027388F: Assay Diluent A, 1x12ml 027388G: Assay Diluent B, 1x12ml 027388H: TMB Substrate, 1x9ml 027388K: Stop Solution, 1x6ml 027388L: Wash Buffer, 30X, 1x20ml Precaution: The Stop Solution (027388K) 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 *027388A, *027388B, *027388D and *027388E 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 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 Substrate Solution. Incubate 15-25 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately.
