Vitamin D Binding Protein (DBP, VDB, Gc-globulin, Group-specific Component, GC), BioAssay(TM) ECL ELISA Kit (Human)

Artikelnummer: USB-158199
Artikelname: Vitamin D Binding Protein (DBP, VDB, Gc-globulin, Group-specific Component, GC), BioAssay(TM) ECL ELISA Kit (Human)
Artikelnummer: USB-158199
Hersteller Artikelnummer: 158199
Alternativnummer: USB-158199-96
Hersteller: US Biological
Kategorie: Kits/Assays
The Human Vitamin D Binding Protein (DBP) ELISA, ECL Kit is a sensitive chemiluminescent sandwich enzyme immunoassay (ECL) for the in vitro quantitative measurement of DBP in human serum, plasma and other biological fluids. Detection Range: 0.47-30ng/ml Sensitivity: < 0.13ng/ml Precision: Intra-Assay: CV<10% Inter-Assay: CV<12% Test Principle: The microtiter strips provided in this kit have been pre-coated with an antibody specific to DBP. A competitive inhibition reaction is launched between a fixed amount of biotin-labeled DBP and unlabeled DBP (in standards or samples) for limited binding sites on the pre-coated antibody. After incubation, the unbound conjugate is washed off. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each well and incubated. Finally, the mixture of substrate A and B is added to generate glow light emission kinetics and the chemiluminescence signal is read using a luminometer. The amount of bound HRP conjugate and the intensity of the emitted light are inversely proportional to the concentration of DBP in the sample or standard. Kit Components: *158199A: Microtiter Plate, 1x96 wells, Pre-coated with antibody, ready to use *158199B: Standard, 2x1vial 158199C: Standard Diluent, 1x20ml *158199D: Detection Reagent A, 1x120ul *158199E: Detection Reagent B, 1x120ul 158199F: Assay Diluent A (2x), 1x6ml 158199G: Assay Diluent B (2x),1x6ml 158199H: Substrate A, 1x10ml 158199K: Substrate B, 1x2ml 158199L: Wash Buffer (30x), 1x20ml Storage and Stability: Store *158199A, *158199B, *158199D and *158199E 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. Materials Required But Not Supplied: 1. Luminometer capable of reading 96-well microplates with the following parameters: lag time 30.0secs, read time 1.0 sec/well. 2. Precision single or multi-channel pipettes and disposable tips. 3. Eppendorf Tubes for diluting samples. 4. Deionized or distilled water. 5. Absorbent paper for blotting the microtiter plate. 6. Container for Wash Solution 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 10 minutes at 37C. 8. Read RLU immediately.