The BioAssay(TM)kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of GRIN2B in rat serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids. Detection Range: 0.312-20ng/ml Sensitivity: 0.117ng/ml Specificity: This assay has high sensitivity and excellent specificity for detection of Glutamate Receptor, Ionotropic, N-Methyl-D-Aspartate 2B (GRIN2B). No significant cross-reactivity or interference between Glutamate Receptor, Ionotropic, N-Methyl-D-Aspartate 2B (GRIN2B) and analogues was observed. Precision: Intra-Assay %CV: <10% Inter-Assay %CV: <12% Test Principle: The microtiter plate provided in this kit has been pre-coated with an antibody specific to GRIN2B. Standards or samples are then added to the appropriate microtiter plate wells with a Horseradish Peroxidase (HRP)-conjugated antibody specific to GRIN2B. After TMB substrate solution is added, those wells that contain GRIN2B will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm 10nm. The concentration of GRIN2B in the samples is then determined by comparing the O.D. of the samples to the standard curve. 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.
