The Human Gastrin (GT) ELISA Kit is a competitive inhibition immunoassay for the in vitro quantitative measurement of GT in human serum, plasma, tissue homogenates, cell lysates, cell culture supernatants and other biological fluids. Detection Range: 12.35-1,000pg/ml Sensitivity: < 4.46pg/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 GT has been pre-coated onto a microplate. A competitive inhibition reaction occurs between biotin-labeled GT and unlabeled GT (standards or samples) with the pre-coated antibody specific for GT. 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 GT in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of GT in the sample. Kit Components: *025250A: Microtiter Plate, 96 wells, Pre-coated, ready to use *025250B: Standard, 2x1vial 025250C: Standard Diluent, 1x20ml *025250D: Detection Reagent A, 1x120ul *025250E: Detection Reagent B, 1x120ul 025250F: Assay Diluent A, 1x12ml 025250G: Assay Diluent B, 1x12ml 025250H: TMB Substrate, 1x9ml 025250K: Stop Solution, 1x6ml 025250L: Wash Buffer, 30X, 1x20ml Storage and Stability: Store *025250A, *025250B, *025250D and *025250E 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.
