The Human HtrA Serine Peptidase (HTRA1) ELISA, ECL Kit is a sandwich Chemiluminescent enzyme immunoassay for the in vitro quantitative measurement of HTRA1 in human serum, plasma and other biological fluids. Test Principle: The microtiter plate provided in this kit has been pre-coated with an antibody specific to HTRA1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to HTRA1. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The addition of substrates A and B mixture generates glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the HTRA1 level in the sample or standard. Detection Range: 8.2 - 6,000pg/ml Sensitivity: <3.2pg/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 HTRA1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to HTRA1. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The addition of substrates A and B mixture generates glow light emission kinetics. Upon plate development, the intensity of the emitted light is proportional to the HTRA1 level in the sample or standard. Kit Components: *157738A: Microtiter Plate, 1x96 wells, Pre-coated, ready to use *157738B: Standard, 2x1vial 157738C: Standard Diluent, 1x20ml *157738D: Detection Reagent A, 1x120ul *157738E: Detection Reagent B, 1x120ul 157738F: Assay Diluent A (2x), 1x6ml 157738G: Assay Diluent B (2x), 1x6ml 157738H: Substrate A, 1x10ml 157738K: Substrate B, 1x2ml 157738L: Wash Buffer (30x), 1x20ml Storage and Stability: Store *157738A, *157738B, *157738D and *157738E 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 100ul Substrate Solution. Incubate 5-10 minutes at 37C. 8. Read RLU immediately.
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