IDK® 25-OH-Vitamin D ELISA

The IDK® 25-OH-Vitamin D ELISA is intended for the quantitative determination of total 25-OH-vitamin D in human serum. For research use only. Not for use in diagnostic procedures.

For Research Use Only in North America. Not For Use in Diagnostic Procedures.

For Laboratory Professional Use Only.

This is the RUO version of Immundiagnostik AG part number K 2020.

Test Principle of the IDK® 25-OH-Vitamin D ELISA

The IDK® 25-OH-Vitamin D ELISA is a sandwich-based enzyme-linked immunosorbent assay. The test employs a very “Exclusive” pair of anti-25-OH Vitamin D Monoclonal Antibodies; the first antibody is immobilized in the solid phase (microtiter wells), and the second antibody is in the liquid phase.

This test will selectively detect 25-OH Vitamin D (including D2 and D3) with high sensitivity and specificity. In the assay procedure, standards and the test specimen (serum) are incubated with the first and second antibodies in the microtiter wells for 20 minutes.

Following a wash step, the resulting Vitamin D-antibody immunocomplex is detected with a third antibody conjugated with horseradish peroxidase (HRP). After a 10-minute incubation, the unbound antibody-HRP conjugate is washed away. Next, a solution of TMB is added for 5–10 minutes of development of the blue color.

The addition of a Stop Solution stops the color development, and the resulting yellow color is measured at 450 nm using a microtiter plate reader. The color intensity will be positively proportional to the concentration of Vitamin D in the sample.

The time for assay completion is 40 minutes.

A dose-response curve of the absorbance unit (optical density, OD at 450 nm) vs. concentration is generated using the values obtained from the standard. The concentration of 25-OH-vitamin D in the samples is determined directly from this curve.