Rat/Mouse SPARC ELISA employs the quantitatively competitive enzyme immunoassay technique in which Rat/Mouse SPARC present in samples competed with a fixed amount of biotinylated Rat/Mouse SPARC for sites on purified rabbit IgG specific against Rat/Mouse SPARC. During the incubation, the standard and samples bound to the Anti SPARC IgG that are pre-coated onto the microplates. The biotinylated SPARC are competitively bound to antibody specific to SPARC. Following a wash to remove any unbound standard, samples and biotin conjugate, a Streptavidin conjugated to horseradish-peroxidase (HRP) is added to the wells. After washing away any unbound enzyme, a substrate solution is added to the wells. The enzyme reaction yields a blue product that turns yellow when the Stop Solution is added. The intensity of the color measured is in proportion to the amount of Rat/Mouse SPARC bound in the initial step. The sample values are then read off the standard curve. Rat/Mouse SPARC ELSA has been shown to accurately quantify the recombinant and natural Rat/Mouse SPARC. Results obtained using natural Rat/Mouse SPARC showed dose response curves that were parallel to the standard curves obtained using the kit standards.
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