RELIABLE:rigorously validated according to FDA/ICH/EMEA guidelines
EASY:ready to use calibrators & controls included
EXCELLENT:correlates with existing methods
DAY TEST:results in 4,5 h
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:
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.
Typical Data
This standard curve and the displayed O.D. values are for demonstration only. A standard curve should be generated for each assay run.
The quality control protocol supplied with the kit shows the results of the final release QC for each kit at the production date. ODs obtained by customers may differ due to various influences including a normal decrease of signal intensity throughout shelf life. However, this does not affect the validity of the results, provided an OD of 1.50 or higher is obtained for the standard with the highest concentration, and the measured control values fall into their respective target range (see labels).
VEGF ELISA Kit Components
All reagents supplied in the human VEGF ELISA kit are stable at 2-8°C until the expiry date stated on the label of each reagent. Reconstituted reagents must be stored at -25°C until the expiry date.
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