FGF-23, Rat, BioAssay(TM) ELISA Kit (Fibroblast Growth Factor 23, Fgf23)

FGF-23, a phosphate-regulating hormone, plays a critical role in phosphorus homeostasis. The study of FGF-23 is important in understanding the role it plays in such processes as bone remodeling and kidney function. FGF-23 is responsible for the inhibition of renal tubular reabsorption of inorganic phosphorus and, consequently, the regulation of phosphorus homeostasis. FGF-23 mRNA is expressed in the brain, in the thymus at low levels, in dendritic cells, with the highest expression in osteocytes and osteoblasts during phases of active bone remodeling. Because circulating levels of FGF-23 are regulated by vitamin D and phosphorus, normal skeletal and soft tissue development require vitamin D and a functional vitamin D signaling pathway. Diseases resulting from aberrant FGF-23 expression include inherited forms of rickets, certain kidney diseases and phosphorus-related tumors\n\nSample Type:\nSerum, plasma, and tissue culture.\n\nIntended Use:\nFor the non-radioactive quantification of Rat FGF-23.\n\nSensitivity:\nThe lowest level of FGF-23 that can be detected by this assay is 0.05ng/ml when using a 50ul sample size.\n\nSpecificity:\nThe antibody pair used in this assay is specific to Rat FGF-23 and does not significantly cross-react to the following molecules/hormones tested: other FGFs\n\nTest Principle:\nThis assay is a Sandwich ELISA based, sequentially, on: 1) concurrent capture of Rat FGF-23 molecules from samples to the wells of a microtiter plate coated with a polyclonal rabbit anti- FGF-23 antibody, and binding of a second biotinylated polyclonal goat anti-FGF-23 antibody to the captured molecules , 2) washing of unbound materials from samples, 3) binding of streptavidin-horseradish peroxidase conjugate to the immobilized biotinylated antibodies, 4) washing of excess of free enzyme conjugates, and 5) quantification of immobilized antibody-enzyme conjugates by monitoring horseradish peroxidase activities in the presence of the substrate 3,3',5,5'- tetramethylbenzidine. The enzyme activity is measured spectrophotometrically by the increased absorbance at 450nm-590nm after acidification of formed products. Since the increase in absorbance is directly proportional to the amount of captured Rat FGF-23 in the unknown sample, the latter can be derived by interpolation from a reference curve generated in the same assay with reference standards of known concentrations of Rat FGF-23.\n\nKit Components:\nFGF-23 ELISA Plate: 1 Strip Plate\nAdhesive Plate Sealer: 2 sheets\nHRP Wash Buffer Concentrate (10X): 2x50ml (Dilute 1:10 with distilled or deionized water)\nFGF-23 Standard (lyophilized): 1x0.25ml upon hydration\nFGF-23 Quality Controls 1 and 2 (lyophilized): 1x0.25ml upon hydration\nMatrix Solution: 1x1.5ml\nAssay Buffer: 1x25ml\nFGF-23 Detection Antibody: 1x12ml\nEnzyme Solution: 1x12ml\nSubstrate (Light sensitive): 1x12ml\nStop Solution (0.3M HCl): 1x12ml\n\nStorage and Stability:\nStore *xxx powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

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