Gamma-Carboxyglutamic Acid (gla) BioAssay(TM) ELISA Kit (General)

Catalog Number: USB-589302
Article Name: Gamma-Carboxyglutamic Acid (gla) BioAssay(TM) ELISA Kit (General)
Biozol Catalog Number: USB-589302
Supplier Catalog Number: 589302
Alternative Catalog Number: USB-589302-96
Manufacturer: US Biological
Category: Kits/Assays
The BioAssay(TM)kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of gla in biological fluids. Detection Range: 1.23-100ng/ml Sensitivity: 0.51ng/ml Specificity: This assay has high sensitivity and excellent specificity for detection of Gamma-Carboxyglutamic Acid (gla). No significant cross-reactivity or interference between Gamma-Carboxyglutamic Acid (gla) and analogues was observed. Precision: Intra-Assay %CV: <10% Inter-Assay %CV: <12% Test Principle: This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to gla has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled gla and unlabeled gla (Standards or samples) with the pre-coated antibody specific to gla. 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 reverse proportional to the concentration of gla in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of gla in the sample. Kit Components: Pre-coated, ready to use 96-well strip plate Standard, 2x vials Detection Reagent A, 1x120ul Detection Reagent B, 1x120ul TMB Substrate, 1x9ml Wash Buffer (30X): 1x20ml Standard Diluent, 1x20ml Assay Diluent A, 1x12ml Assay Diluent B, 1x12ml Stop Solution, 1x6ml Storage and Stability: Store Microtiter Strips, Standard, Detection Reagents and Positive Control at -20C. After reconstitution of Positive Control, store at 4C and use within 5 days. Store all the other components at 4C. Unused kit is stable for 6 months after receipt. 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 1 hour 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 90ul Substrate Solution. Incubate 10-20 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately.