The gene for human VEGF is organized into 8 exons. As a result of alternative splicing, at least 4 transcripts encoding mature monomeric VEGF containing 121, 165, 189, and 206aa residues (VEGF121, VEGF165, VEGF189, and VEGF206). VEGF is a highly conserved protein that has cross-species activity. Between human, rat or bovine VEGF, 84-94% sequence identity has been observed. In vitro, VEGF is a potent endothelial cell mitogen. In cultured endothelial cells, VEGF can activate phospholipase C and induce rapid increases of free cytosolic Ca2+. VEGF has also been shown to be chemotactic for monocytes and osteoblasts. Based on its in vitro and in vivo properties, VEGF is expected to play important roles in inflammation and during normal and pathological angiogenesis, a process that is associated with wound healing, embryonic development, and growth and metastasis of solid tumors.
Sample Type:Cell culture supernatants, serum, plasma, and tissue
Intended Use:The Human VEGF BioAssay™ ELISA Kit is an enzyme-linked immunosorbent assay for the quantitative detection of Human VEGF concentrations in cell culture supernatants, serum, plasma, and tissue.
Sensitivity:4pg/ml
Range:15.625-1000pg/ml
Specificity:Recognizes human VEGF. There was no detectable cross reactivity with any of the tested proteins following IL-1,2,3,4,5,6, 8,10,12,TNF,TGF and IFN.
Sample Volume:100ul/well
Test Principle:This assay employs the Sandwich immunoassay technique. An anti-h VEGF monoclonal coating antibody is adsorbed onto microwells. VEGF present in the sample or standard binds to antibodies adsorbed to the microwells. Following incubation unbound sample or standard are removed during a wash step. a Biotinylated anti-h VEGF antibody is added and binds to VEGF captured by the first antibody. Following incubation unbound Biotinylated anti-h VEGF antibody is removed during a wash step. A Streptavidin-HRP is added and binds to Biotinylated anti-h VEGF antibody. Following incubation unbound Streptavidin-HRP is removed during a wash step. A colored product is formed in proportion to the amount of VEGF present in the sample. The reaction is terminated by addition of acid and absorbance is measured at 450nm. A standard curve is prepared from seven VEGF standard dilutions and VEGF sample concentration determined.
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
