Bovine Vascular Endothelial Growth Factor 165 ELISA Kit

A sandwich enzyme-linked immunosorbent assay from Antibodies.com, this kit is designed for the in vitro quantitative determination of bovine VEGF165 in serum, plasma, tissue homogenates, cell lysates, cell culture supernates, and other biological fluids.

The assay employs a sandwich immunoassay technique using a pre-coated microplate. Standards and test samples are added to the wells, where VEGF165 is captured by an immobilized antibody. Following incubation and washing, a Biotinylated Anti-VEGF165 Antibody is added and binds to the captured VEGF165. HRP-Avidin conjugate is then added, and after further incubation and washing, TMB substrate solution is used to visualize the enzymatic reaction. The reaction produces a blue-coloured product that changes to yellow upon addition of acidic stop solution. The intensity of yellow is proportional to the amount of VEGF165 captured, and concentration is calculated by reading absorbance at 450 nm and referring to a standard curve.

The assay has a quantitative measuring range of 31.2-2000 pg/ml and a sensitivity of <8.3 pg/ml. It is intended for use with a colorimetric microplate reader, and the total assay time is 4h 30m. Recovery was evaluated in serum (range 83-100%, average 91%, n=5), EDTA plasma (range 90-98%, average 94%, n=5), and heparin plasma (range 89-102%, average 95%, n=5). Dilution linearity was demonstrated at 1:2, 1:4, 1:8, and 1:16 dilutions for serum, EDTA plasma, and heparin plasma samples (n=5 each). The kit includes a pre-coated microplate, lyophilized standard, detection solutions A and B, wash buffer, sample dilution buffer, TMB substrate, stop solution, and plate sealers. Components are stored at +4°C or -20°C as instructed, and the kit should not be used past its expiration date.