Anti-Phospho-VEGFR2/KDR (Tyr1059) Polyclonal Antibody is a Rabbit antibody targeting Phospho-VEGFR2/KDR (Tyr1059). Anti-Phospho-VEGFR2/KDR (Tyr1059) Polyclonal Antibody can be used in FCM,IF,IHC-Fr,IHC-P,WB.

IgG

Mouse,Human (predicted:Rat,Chicken,Dog,Pig,Cow,Horse,Rabbit)

1. Tissue/cell: human kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-VEGFR Polyclonal Antibody, Unconjugated (TMAB-01528) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Blank control: Huvec.
Primary Antibody (green line): Rabbit Anti-phospho-VEGF receptor 2 (Tyr1059) antibody (TMAB-01528)
Dilution: 2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. Sample:
HUVEC (Hu) lysate at 30 μg
Primary: Anti-phospho-VEGF receptor 2 (Tyr1059) (TMAB-01528) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 147 kDa
Observed band size: 147 kDa

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 2ug/test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human VEGF receptor 2 around the phosphorylation site of Tyr1059

Human

KDR

Vascular endothelial growth factor receptor 2

Response to hypoxia,VEGF,Angiogenic growth factors,Growth factor receptors,Endothelial Cell Markers,VEGF Receptors,Endothelial Cells,Hypoxia,Host Virus Interaction,VEGF,Receptor Tyrosine Kinases,Endothelial Markers,Surface Molecules

Tyrosine-protein kinase that acts as a cell-surface receptor for VEGFA, VEGFC and VEGFD. Plays an essential role in the regulation of angiogenesis, vascular development, vascular permeability, and embryonic hematopoiesis. Promotes proliferation, survival, migration and differentiation of endothelial cells. Promotes reorganization of the actin cytoskeleton. Isoforms lacking a transmembrane domain, such as isoform 2 and isoform 3, may function as decoy receptors for VEGFA, VEGFC and/or VEGFD. Isoform 2 plays an important role as negative regulator of VEGFA-and VEGFC-mediated lymphangiogenesis by limiting the amount of free VEGFA and/or VEGFC and preventing their binding to FLT4. Modulates FLT1 and FLT4 signaling by forming heterodimers. Binding of vascular growth factors to isoform 1 leads to the activation of several signaling cascades. Activation of PLCG1 leads to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate and the activation of protein kinase C. Mediates activation of MAPK1/ERK2, MAPK3/ERK1 and the MAP kinase signaling pathway, as well as of the AKT1 signaling pathway. Mediates phosphorylation of PIK3R1, the regulatory subunit of phosphatidylinositol 3-kinase, reorganization of the actin cytoskeleton and activation of PTK2/FAK1. Required for VEGFA-mediated induction of NOS2 and NOS3, leading to the production of the signaling molecule nitric oxide (NO) by endothelial cells. Phosphorylates PLCG1. Promotes phosphorylation of FYN, NCK1, NOS3, PIK3R1, PTK2/FAK1 and SRC.