The Antidiuretic Hormone (ADH) BioAssay™ ECL ELISA Kit (Human) is a sensitive chemiluminescent competitive enzyme immunoassay for the in vitro quantitative measurement of ADH in human serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids.
Detection Range:11.72-3000pg/ml
Sensitivity:<4.66pg/ml
Precision:Intra-Assay: CV<10%Inter-Assay: CV<12%
Test Principle:The microtiter plate provided in this kit has been pre-coated with a monoclonal antibody specific to ADH. Competitive binding is launched between a fixed amount of biotin-labeled ADH and unlabeled ADH (in the standard or sample) for limited binding sites on the pre-coated antibody. After incubation, unbound materials are washed off. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and the plate is incubated. The addition of subtrate A and substrate B mixture generates glow light emission kinetics. The intensity of the emitted light is inversely proportional to the concentration of ADH in the sample or standard.
Kit Components:*365717A: Microtiter Plate, 1x96 wells, Pre-coated with antibody; ready to use
*365717B: Standard, 2x1vial 365717C: Standard Diluent, 1x20ml*365717D: Detection Reagent A, 1x120ul *365717E: Detection Reagent B, 1x120ul 365717F: Assay Diluent A (2x), 1x12ml365717G: Assay Diluent B (2x), 1x12ml365717H: Substrate A, 1x10ml365717K: Substrate B, 1x2ml365717L: Wash Buffer (30x), 1x20ml
Storage and Stability:Store *365717A, *365717B, *365717D and *365717E at -20°C. Store all the other components at 4°C. 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 50ul standard or sample to each well, then add 50ul Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37°C
3. Aspirate and wash 3 times
4. Add 100ul Detection Reagent B. Incubate 30 minutes at 37°C
6. Aspirate and wash 5 times
7. Add 100ul Substrate Solution. Incubate 10 minutes at 37°C
