Description
The Angiopoietin-2 human ELISA kit is a sandwich enzyme immunoassay for the quantitative determinationof angiopoietin-2 in human serum and plasma samples. Standards, controls and samples must be pre-diluted 1+10 prior to assaying. In a first step, pre-diluted standard/control/sample and biotinylated antibody (goat polyclonal anti-human angiopoietin-2) are pipetted into the wells of the microtiter strips, which are pre-coated with a monoclonal anti-human angiopoietin-2 antibody. Angiopoietin-2 present in the standard/control/sample binds to the pre-coated antibody in the well and forms a sandwich with the anti-human angiopoietin-2 antibody. In a washing step all non-specific unbound material is removed. In a second step, the conjugate (Streptavidin-HRP) is pipetted into the wells and reacts with the biotinylated antibody. After another washing 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 angiopoietin-2 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) using the values obtained from the standards versus the standard concentration is generated. The concentration of angiopoietin-2 in the sample is determined directly from the dose response curve