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The FGF-23 ELISA Kit is a two-site enzyme-linked immunosorbent assay kit for the measurement of FGF-23 in serum. Two specific murine monoclonal antibodies bind to full-length FGF-23. One antibody is immobilized onto the microtiter plate well for capture. The other antibody is conjugated to HRP(horseradish peroxidase) for detection.
In first reaction, a sample containing FGF-23 is incubated with the immobilized antibody in a microtiter well. FGF-23 in the sample is captured with the antibody. At the end of this reaction, the well is washed to remove unbound FGF-23 and other components.
In second reaction, this immobilized FGF-23 is incubated with HRP labeled antibody to form a “sandwich”complex :
Anti FGF-23 antibody - N-terminal FGF-23 C-terminal - HRP labeled anti FGF-23 antibody
At the end of this reaction, the well is washed to remove unbound components.
In enzyme reaction, the sandwich complex immobilized on the well is incubated with a substrate solution and then measured by a spectrophotometric microtiter plate reader. The enzymatic activity of the complex bound to the well is directly proportional to the amount of FGF-23 in the sample.
A standard curve is generated by plotting the absorbance versus the each concentration of FGF-23 standard. The concentration of FGF-23 in the sample is determined from this curve. |
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