Description
This Eagle Biosciences High Sensitive Human Fetuin-A ELISA Assay Kit is designed, developed and produced for the quantitative measurement of human Fetuin-A in serum samples. The assay utilizes the two-site “sandwich” technique with two selected goat anti-human Fetuin-A polyclonal antibodies that bind to different epitopes of human Fetuin-A. Assay standards, controls and prediluted patient serum samples containing human Fetuin-A are added to microtiter wells of a microplate that was coated with a high affinity polyclonal goat anti-human Fetuin-A antibody. After the first incubation period, the antibody on the wall of microtiter well captures human Fetuin-A in the sample and unbound proteins in each microtiter well are washed away. Then a horseradish peroxidase (HRP)-conjugated polyclonal anti-human Fetuin-A antibody is added to each microtiter well and a “sandwich” of “capture antibody – human Fetuin-A – HRP-conjugated tracer antibody” is formed. The unbound tracer antibody is removed in the subsequent washing step. HRP-conjugated tracer antibody bound to the well is then incubated with a substrate solution in a timed reaction and then measured in a spectrophotometric microplate reader. The enzymatic activity of the tracer antibody bound to the Fetuin-A on the wall of the microtiter well is directly proportional to the amount of Fetuin-A in the sample. A standard curve is generated by plotting the absorbance versus the respective human Fetuin-A concentration for each standard on point-to-point or cubical scales. The concentration of human Fetuin-A in test samples is determined directly from this standard curve