Product Description and Product Data
This is an all-inclusive cell-based luciferase reporter assay kit targeting the Human Vascular Endothelial Growth Factor Receptor 2 (VEGFR2). INDIGO’s VEGFR2 reporter assay utilizes proprietary mammalian cells that have been engineered to provide constitutive expression of VEGFR2. In addition to VEGFR2 Reporter Cells, this kit provides two optimized media for use during cell culture and in diluting the user’s test samples, a reference agonist, Luciferase Detection Reagent, and a cell culture-ready assay plate. The principal application of this assay is in the screening of test samples to quantify any functional activity, either agonist or antagonist, that they may exert against human VEGFR2. This kit provides researchers with clear, reproducible results, exceptional cell viability post-thaw, and consistent results lot to lot. Kits must be stored at -80C. Do not store in liquid nitrogen. Note: reporter cells cannot be refrozen or maintained in extended culture.
Clear, Reproducible Results
- All-Inclusive Assay Systems
- Exceptional Cell Viability Post-Thaw
- Consistent Results Lot to Lot
|Target Type||Growth Factor Receptor|
|Shelf Life||6 months|
|Shipping Requirements||Dry Ice|
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.
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