Biotinidase (BTD) BioAssay(TM) ELISA Kit (Human)

The Human Biotinidase (BTD) ELISA Kit is a competitive inhibition immunoassay for the in vitro quantitative measurement of BTD in human serum, plasma and other biological fluids. Detection Range: 123.5 � 10,000pg/ml Sensitivity: 42.6pg/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 BTD has been pre-coated onto a microplate. A competitive inhibition reaction occurs between biotin-labeled BTD and unlabeled BTD (standards or samples) with the pre-coated antibody specific for BTD. 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 BTD in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of BTD in the sample. Kit Components: *168001A: Microtiter Plate, 96 wells, Pre-coated, ready to use *168001B: Standard, 2x1vial 168001C: Standard Diluent, 1x20ml *168001D: Detection Reagent A, 1x120ul *168001E: Detection Reagent B, 1x120ul 168001F: Assay Diluent A, 1x12ml 168001G: Assay Diluent B, 1x12ml 168001H: TMB Substrate, 1x9ml 168001K: Stop Solution, 1x6ml 168001L: Wash Buffer, 30X, 1x20ml Storage and Stability: Store *168001A, *168001B, *168001D and *168001E at -20 degrees C. Store all the other components at 4 degrees 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 prepared Detection Reagent A immediately. Shake and mix. Incubate 1 hour at 37 degrees C. 3. Aspirate and wash 3 times. 4. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37 degrees C. 5. Aspirate and wash 5 times. 6. Add 90ul Substrate Solution. Incubate 15-25 minutes at 37 degrees C. 7. Add 50ul Stop Solution. Read absorbance at 450nm immediately.