Human VEGF ELISA Product Overview

The Biomedica human Vascular Endothelial Growth Factor (VEGF) ELISA kit is a 4.5 hour 96-well  sandwich enzyme immunoassay for the quantitative determination of human VEGF in serum, plasma (EDTA, citrate) , cell culture supernatants, and urine samples.

The VEGF ELISA assay recognizes both natural and recombinant human VEGF with high sensitivity. The assay employs highly specific epitope mapped antibodies as well as eight human serum-based standards and two controls to ensure the measurement of biologically reliable data.

Human VEGF ELISA Assay Principle

The human Vascular Endothelial Growth Factor (VEGF) ELISA kit is a sandwich enzyme immunoassay that has been optimized and fully validated for the quantitative determination of human VEGF in serum, EDTA-plasma, and citrate plasma.  Validation experiments have been performed according to international quality guidelines (ICH/ FDA/ EMEA).  Cell culture supernatant and urine samples are compatible with this ELISA. The VEGF ELISA assay recognizes both natural and recombinant human VEGF.

The figure below explains the principle of the human VEGF sandwich ELISA:

Capture antibody: recombinant anti-human VEGF antibody
Detection antibody: polyclonal anti-human VEGF antibody, Biotin-labeled
Target antigen: human VEGF

This kit is a sandwich enzyme immunoassay for the quantitative determination of human VEGF (Vascular Endothelial Growth Factor) in human serum, plasma, cell culture supernatants, and urine samples.  In a first step, assay buffer is pipetted into the wells of the microtiter strips. Thereafter, STD/sample/CTRL are pipetted into the wells, which are pre-coated with the recombinant anti-human VEGF antibody.  Any soluble VEGF present in the STD/sample/CTRL binds to the pre-coated anti-VEGF antibody in the well. After incubation, a washing step is applied where all non-specific unbound material is removed. In a next step, the biotinylated anti-VEGF antibody (AB) is pipetted into the wells and reacts with the VEGF present in the sample, forming a sandwich. Next, all unbound antibody is removed during another washing step. In the next step, the conjugate (streptavidin-HRPO) is added and reacts with the biotinylated anti-VEGF antibody. After another washing step, the substrate (Tetramethylbenzidine; TMB) is pipetted into the wells. The enzyme catalysed colour change of the substrate is directly proportional to the amount of VEGF present in the sample. 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) versus standard concentration is generated, using the values obtained from the standards. The concentration of soluble VEGF in the sample is determined directly from the dose response curve.