Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF), a potent cytokine expressed by most malignant tumors, has critical roles in vasculogenesis and both physiological and pathological angiogenesis.VEGF produced by tumor cells potently stimulates endothelial cell proliferation and angiogenesis and plays a key role in the pathophysiology of several neoplasias. VEGF may also play a pivotal role in mediating the development and progression of diabetic retinopathy.VEGF, a major regulator of angiogenesis, binds to two receptor tyrosine kinases, KDR/Flk-1 and Flt-1. The VEGF gene is mapped by fluorescence in situ hybridization to chromosome 6p12.
Intended Use:VEGF BioAssay™ ELISA Kit, Human is a sandwich ELISA for quantitative detection of human VEGF in cell culture supernates, serum and plasma (heparin, EDTA, citrate).
Storage and Stability:Store *144095B powder at 4°C. Once reconstituted store at 4°C for up to 12 hours or at -20°C for up to 48 hours. Store other components at 4°C. Stable for 6 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
Assay Summary:1. Add samples and standards and incubate the plate at 37°C for 90 minutes. Do not wash. 2. Add biotinylated antibody and incubate the plate at 37°C for 60 minutes. Wash plate 3 times with 0.01M PBS or TBS
3. Add ABC working solution and incubate the plate at 37°C for 30 minutes. Wash plate 5 times with 0.01M PBS or TBS
4. Add TMB color developing agent and incubate the plate at 37°C in dark for 20-25 minutes
