The Insulin ELISA Assay Kit is intended for use in the quantitative determination of human insulin in serum and/or plasma. This Insulin ELISA Assay Kit is designed, developed, and produced for the quantitative measurement of human insulin in serum and /or EDTA-plasma samples.
The assay uses the "sandwich" technique with selected antibodies that bind to different insulin epitopes. You can easily get the human insulin ELISA online.
Standards for analysis, control, and patient samples were added directly to the bore of the microtiter plate, which was coated with an anti-human insulin-specific antibody. At the same time, a monoclonal insulin-specific antibody conjugated with horseradish peroxidase was added to each well.
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After the first incubation period, antibodies on the walls of the microtiter wells captured human insulin in the samples, and the unbound protein in each microtiter well was washed off. A "sandwich" is formed of "anti-insulin antibody – human insulin – HRP conjugate tracking antibody".
The next unbound antibody is removed in the next washing step. To detect these immune complexes, the wells were then incubated with the substrate solution in a specific reaction and then measured in a spectrophotometric microplate reader.
The enzyme activity of the immune complex bound to human insulin on the microtiter well wall is directly proportional to the amount of insulin in the sample.
Standard curves were created by plotting the absorbance relative to the corresponding human insulin concentration for each standard on a point-to-point or 4-parameter curve. The concentration of human insulin in the test sample is determined directly from this standard curve.