Total-VEGFR2 cellular kit HTRF®
The Total VEGFR2 kit monitors the cellular VEGFR2 expression level and can be used as a normalization assay for the Phospho-VEGFR2 kit.
- High sensitivity
- Faster and more convenient than ELISA
Homogeneous cell-based assay for monitoring total VEGFR2 modulations. VEGFR2 mediated signaling data has uses in angiogenesis, vascular permeability, and cancer stem cell regulation research.
- VALIDATED ON MULTIPLE CELL TYPES
Huvec cells were plated at 150,000 cells per well in a 96-well plate. After an overnight incubation at 37°C, 5% CO2, a serial dilution of human VEGF was added to the cells for 3 minutes at 37°C, 5% CO2. Stimulation medium was removed from cells and 50µL of lysis buffer were added. A lysis step was carried out, shaking gently for 30 minutes. 16µL of samples were transferred into a 384-well small volume plate, then 4µL of Phospho VEGFR2 HTRF detection reagents were added. In parallel, 16µl were dispensed into other wells, then 4µL of HTRF Total VEGFR2 detection reagents were added. Signals were recorded overnight.
Human Huvec cells were cultured to 80% confluency. After 3 minutes of hVEGF treatment, cells were lysed and soluble supernatants were collected via centrifugation. Serial dilutions of the cell lysate were performed, and 16 µL of each dilution were transferred into a 384-well low volume white microplate before finally adding VEGFR2 cellular kit detection reagents.
A side by side comparison showed the HTRF total assay is at least 32-fold more sensitive than the Western Blot.
The vascular endothelial growth factor (VEGF) is an endothelial-specific mitogen factor that is often associated with the activation of tumour-induced angiogenesis. In tumours, VEGF secretion is upregulated. At the endothelial cell surface, the secreted protein binds with high affinity to at least two distinct vascular endothelial growth factor receptors (VEGFRs), VEGFR-1, also known as fms-like tyrosine kinase (Flt-1) and VEGFR-2, also known as fetal liver kinase (Flk-1).
Activation of endothelial cells by VEGF leads to the receptor auto-phosphorylation and subsequent signaling. Multiple phosphorylation sites have been described.
VEGFR is implicated in angiogenesis, and
combined inhibition of VEGF and PDGF signaling enforces tumor vessel regression.
Physiologically relevant results fo fast flowing research - Flyers
Insider Tips for successful sample treatment - Technical Notes
HTRF and WB compatible guidelines - Technical Notes
Protocol for tumor xenograft analysis with HTRF - Technical Notes
Mastering the art of cell signaling assays optimization - Guides
Multi-tissue cellular modeling and anlysis of insulin signaling - Posters
A solution for phospho-protein analysis in metabolic disorders - Posters
Detailed protocol and direct comparison with WB - Posters
A single technology for 2D cells, 3D cells, and xenograft models - Posters
PI3K/AKT/mTor translational control pathway - Posters
Analysis of a large panel of diverse biological samples and cellular models - Posters
One technology across all samples - Application Notes
Tumor xenograft analysis: HTRF versus Western blot - Application Notes
Valuable guidelines for efficiently analyzing and interpreting results - Application Notes
Increased flexibility of phospho-assays - Application Notes
Analyse of PI3K/AKT/mTor translational control pathway - Application Notes
In collaboration with Bayer - Scientific Presentations
A fun video introducing you to phosphorylation assays with HTRF - Videos
64VEGFR2TPEG-64VEGFR2TPEH - Product Insert
Seeding and lysing recommendations for a number of cell culture vessels. - Technical Notes
Combination of AlphaLISA®, HTRF®, or AlphaLISA® SureFire® Ultra™ immunoassays with the ATPlite™ 1step cell viability assay - Application Notes
Learn how to reduce time and sample consumption - Application Notes
Plate Reader Requirement
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