Description
This kit is a sandwich enzyme immunoassay for the direct determination of total soluble Neuropilin-1 in humanserum and plasma samples. STD/CTRL/Sample require pre-treatment with an equal amount of guanidine hydrochloride (GuHCl) to remove potentially bound ligands, followed by pre-dilution with assay buffer. In a next step, pre-treated and diluted STD/CTRL/Sample and detection antibody (mouse monoclonal anti human Neuropilin-1 IgG) are pipetted into the wells of the microtiter strips, which are pre-coated with anti Neuropilin-1 antibody. Neuropilin-1 present in STD/CTRL/Sample binds to the pre-coated antibody in the well and forms a sandwich with the detection antibody. In the washing step all non-specific unbound material is removed. Subsequently, the conjugate (Streptavidin-HRPO) is pipetted into the wells and reacts with the detection antibody. After another washing step, the substrate (TMB Tetramethylbenzidine) is pipetted into the wells. The enzyme catalysed colour change of the substrate is directly proportional to the amount of Neuropilin-1. This colour change is detectable with a standard microtiter plate ELISA reader. A dose response curve of the absorbance (optical density, OD at 450 nm) vs. standard concentration is generated, using the values obtained from the standard. The concentration of Neuropilin-1 in the sample is determined directly from the dose response curve