Vanin-1 ELISA

The Vanin-1 (urine) ELISA kit is a sandwich enzyme immunoassay for the quantitative determination of Vanin-1 in human urine samples. In a first step, assay buffer is pipetted into the wells of the microtiter strips. Thereafter, standard/control/sample and detection antibody (CONJ, polyclonal sheep anti-human Vanin-1-HRP) are pipetted into the wells, which are precoated with polyclonal sheep anti-human Vanin-1 antibody. Vanin-1 present in the standard/control/sample binds to the pre-coated antibody in the well and forms a sandwich with the detection antibody. In the washing step, all nonspecific unbound material is removed. In a next step, the substrate (TMB, tetramethylbenzidine) is pipetted into the wells. The enzyme-catalyzed color change of the substrate is directly proportional to the amount of Vanin-1 present in the sample. This color change is detectable with a standard microplate reader. A dose response curve of the absorbance (optical density, OD at 450 nm) versus standard concentration is generated using the values obtained from the standards. The concentration of Vanin-1 in the sample is determined directly from the dose response curve.