The Thyroxine (T4) ELISA Kit is a competitive inhibition immunoassay for the in vitro quantitative measurement of T4 in serum, plasma and other biological fluids. Detection Range: 3.70-300ng/ml Sensitivity: <1.36ng/ml Precision: Intra-Assay CV: <10% Inter-Assay CV: <12% Test Principle: This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific for T4 has been pre-coated onto a microplate. A competitive inhibition reaction occurs between biotin-labeled T4 and unlabeled T4 (standards or samples) with the pre-coated antibody specific for T4. After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is inversely proportional to the concentration of T4 in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of T4 in the sample. Kit Components: *028408A: Microtiter Plate, 96 wells, Pre-coated, ready to use *028408B: Standard, 2x1vial 028408C: Standard Diluent, 1x20ml *028408D: Detection Reagent A, 1x1vial *028408E: Detection Reagent B, 1x120ul 028408F: Assay Diluent A, 1x12ml 028408G: Assay Diluent B, 1x12ml 028408H: TMB Substrate, 1x9ml 028408K: Stop Solution, 1x6ml 028408L: Wash Buffer, 30X, 1x20ml 028408M: Reagent Diluent, 1x300ul Storage and Stability: Store *028408A, *028408B, *028408D and *028408E 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 50ul standard or sample to each well, then add 50ul prepared Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37C. 3. Aspirate and wash 3 times. 4. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37C. 5. Aspirate and wash 5 times. 6. Add 90ul Substrate Solution. Incubate 15-25 minutes at 37C. 7. Add 50ul Stop Solution. Read absorbance at 450nm immediately.
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