| Podoplanin (PDPN) BioAssay(TM) ELISA Kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of PDPN in human tissue homogenates, cell lysates and other biological fluids. Detection Range: 0.156-10ng/ml Sensitivity: 0.056ng/ml Intra-Assay: CV<10% Inter-Assay: CV<12% Kit Components: *027544A: Microtiter Strips, 1x96 wells, Pre-coated, ready to use *027544B: Standard, 2x1 vial 027544C: Standard Diluent, 1x20ml *027544D: Detection Reagent A (green), 1x120ul *027544E: Detection Reagent B (red), 1x120ul 027544F: Assay Diluent A, 1x12ml 027544G: Assay Diluent B, 1x12ml 027544H: TMB Substrate, 1x9ml *027544J: Positive Control, 5.28ng/ml, 1x1vial (Lyophilized) 027544K: Stop Solution, 1x6ml 027544L: Wash Buffer, 30X, 1x20ml Precaution: The Stop Solution (027544K) included for use in this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. Storage and Stability: Store *027544A, *027544B, *027544D, *027544E and *027544J at -20C. After reconstitution of *027544J, store at 4C and use within 5 days. 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. Notes on 027544J: Positive Control Intended Use: The positive control is from human U87-MG cell lysates known to contain the target protein. It will provide a positive result in the ELISA assay, which indicates that the kit is optimized and working properly. Characteristics: The positive control vial contains human PDPN in human serum (lyophilized powder). Note: The human based material used in this product is screened for HIV, Hepatitis A, B, C, HTLV and Syphilis. Expected Values: 5.28ng/ml Usage: Dissolve with 150ul standard diluent and let stand for 10 minutes. If powder is partly insoluble, shake gently until fully dissolved. Mix gently before use. In the experiment, add 100ul to an appropriate well of the plate and test its concentration along with standards and your specimens. Note: Upon receipt store positive control vial at -20C. After reconstitution, store Positive Control vial at 4C and use within 5 days. Assay Procedure Summary: 1. Prepare all reagents, samples and standards. 2. Add 100ul standard or sample to each well. Incubate 1 hour 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 10-20 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately. |