Vascular Endothelial Growth Factor Receptor 2 (VEGFR2)
|Product Family||Product Number||Product Description||Technical Manual|
|IB15001-32||Human VEGFR2 Reporter Assay System, 3 x 32 assays in 96-well format||Technical Manual|
|IB15001||Human VEGFR2 Reporter Assay System, 1 x 96-well format assays||Technical Manual|
|IB15002||Human VEGFR2 Reporter Assay System, 1 x 384-well format assays|
Human Vascular Endothelial Growth Factor Receptor 2 Assay Kit
This Human type II Vascular Endothelial Growth Factor Receptor (VEGFR2) kit product is an all-inclusive assay system that includes addition to VEGFR2 Reporter Cells, two optimized media for use in recovering the cryopreserved cells and for diluting test samples, the reference activator human VEGF-A (165 aa isoform, non-glycosylated), Luciferase Detection Reagents, and a cell culture-ready assay plate.
VEGFR2 Reporter Cells are transiently transfected and prepared as frozen stocks using INDIGO's proprietary CryoMite™ process. This cryo-preservation method allows for the immediate dispensing of healthy, division-competent reporter cells into assay plates. There is no need for cumbersome intermediate treatment steps such as spin-and-rinse of cells, viability determinations, or cell titer adjustments prior to assay setup.
INDIGO’s VEGFR2 assay kits feature a luciferase detection reagent specially formulated to provide stable light emission between 5 and 90+ minutes after initiating the luciferase reaction. Incorporating a 5-minute reaction-rest period ensures that light emission profiles attain maximal stability, thereby allowing assay plates to be processed in batch. By doing so, the signal output from all sample wells, from one plate to the next, may be directly compared within an experimental set.
VEGFR2 reporter assay kits are offered in different assay formats to accommodate researchers' needs: 3x 32, 1x 96, and 1x 384 assay formats for screening small numbers of test compounds, as well as custom bulk reagents for HTS applications.
Bulk assay reagents can be custom manufactured to accommodate any scale of HTS. Please inquire.
Human Vascular Endothelial Growth Factor Receptor 2 Assay Services
The primary application of INDIGO's cell-based VEGFR2 assays are to quantitatively assess the bioactivity of a test compound as an agonist or antagonist of the receptor. Service assays include a positive control reference compound and 'vehicle' control for every experiment. A formal study report and all data files are provided to the client upon completion of the study. To receive a quote for your proposed study, complete & submit the online "Request a Quote" form or contact an INDIGO Customer Service Representative to discuss your desired study parameters.
Human Vascular Endothelial Growth Factor Receptor 2 Assay Background
There are several isoforms of the physiological signaling molecule vascular endothelial growth factor (VEGF); the 165aa isoform of VEGF-A is the reference activator provided with this kit. Stated simplistically, VEGF-A forms a complex with heparin (or heparan sulphate) and other co-receptors, which then bind to the extra-cellular domain of vascular endothelial growth factor receptor type II (VEGFR2). Dimerization, activation of cytoplasmic tyrosine kinase domains and auto-phosphorylation ensue, followed by phosphorylation and activation of alternative associated intracellular signaling proteins.
VEGFR2, a member of the receptor tyrosine kinase (RTK) family, is highly expressed in vascular endothelial cells. VEGFR2 is a master regulator of the normal physiological processes of vasculogenesis, angiogenesis and lymphangiogenesis. Alternatively, dysregulated expression of VEGFR2 is strongly associated with cancer progression and tumor development.
Activated VEGFR2 may signal through several different pathways, the most prominent being Src, GRB2/Shc/SOS, PI3K, PLC-γ and JAK/STAT. These various signal transduction pathways culminate in the activation of pathway-related transcription factors and the induced expression of their respective target genes.
Phosphorylation and activation of the transcription factor NFAT is one outcome of VEGFR2 activation, and it is the signaling mechanism exploited by the reporter cells included in this kit. Accordingly, INDIGO's Reporter Cells contains an engineered luciferase reporter gene functionally linked to tandem NFAT genetic response element (GRE) sequences and a minimal promoter. Activated NFAT will bind to its corresponding GRE’s to initiate the formation of a complete transcription complex that drives Luc gene expression. Quantifying relative changes in luciferase activity in the treated reporter cells relative to the untreated cells provides a sensitive, dose-dependent surrogate measure of drug- or antibody-induced changes in VEGFR2 activity.
Considering the significant involvement of VEGFR2 in sustaining tumor development it continues to command much interest as a target for the development of novel, specific small molecule inhibitory drugs and antibodies. Accordingly, the primary application of this reporter assay is to screen test materials for any functional activity, either agonistic or inhibitory, that they may exert against VEGFR2 or the activator protein VEGF-A.
This assay utilizes proprietary human cells that provide constitutive expression of the Human type II Vascular Endothelial Growth Factor Receptor (VEGFR2), also known as KDR and FLK-1.