| This Fibroblast Growth Factor 23 (FGF23) BioAssay(TM) ELISA Kit is a sandwich ELISA for in vitro quantitative measurement of FGF23 in mouse serum, plasma and other biological fluids. Detection Range: 15.63-1000pg/ml Sensitivity: 9.38pg/ml Precision: Intra-Assay %CV: <10% Inter-Assay %CV: <10% Specificity: Recognizes mouse FGF23. No significant crossreactivity or interference between mouse FGF23. and analogues was observed. Test Principle: The microtiter plate provided in this kit has been pre-coated with an antibody specific to mouse FGF23. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to FGF23. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Free components are washed away. The substrate solution is added, only those wells that contain FGF23, biotin-conjugated antibody and enzyme-conjugated Avidin will apear blue in color. The enzyme-substrate reaction is terminated by the addition of sulfuric acid solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450nm 2nm. The OD value is proportional to the concentration of mouse FGF23. The concentration of FGF23 in the sample is then determined by comparing the O.D. of the sample to the standard curve. Kit Components: *382872A: Microtiter Strips, 1x96 wells (8x12wells), Pre-coated, ready to use *382872B: Standard, 2x1vial 382872C: Standard Diluent, 1x20ml *382872D: Biotin Detection, 100X, 1x120ul *382872E: HRP Conjugate, 100X, 1x120ul 382872F: Biotin Detection Diluent, 1x14ml 382872G: HRP Diluent, 1x14ml 382872H: Substrate, 1x10ml 382872K: Stop Solution, 1x10ml 382872L: Wash Buffer, 25X, 1x30ml Precaution: The Stop Solution (382872K) included for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material. Storage and Stability: Store *382872A, *382872B, *382872D and *382872E at -20C. 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 90 minutes at 37C. 3. Aspirate and add 100ul prepared Biotin Working Solution. Incubate 1 hour at 37C. 4. Aspirate and wash 3 times. 5. Add 100ul preparedHRP Detection Working Solution. Incubate 30 minutes at 37C. 6. Aspirate and wash 5 times. 7. Add 90ul Substrate. Incubate 15 minutes at 37C. 8. Add 50ul Stop Solution. Read at 450nm immediately. |
