Mouse, Human Predicted to work with:
Synthetic phospho peptide containing Tyrosine 1054.
The sequence is conserved in mouse and rat.
NIH 3T3 cells transfected with full-length, wild type VEGFR 2, or Porcine Aortic cells transfected with a chimeric receptor consisting of the extracellular domain receptor coupled to the transmembrane and cytoplasmic domains of the VEGFR 2.
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Purification notesThe antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated VEGFR 2 protein. The final product is generated by affinity chromatography using a VEGFR 2 derived peptide that is phosphorylated at Tyrosine 1054.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 0.35 - 1 µg/ml. Detects a band of approximately 200 kDa.
FunctionReceptor for VEGF or VEGFC. Has a tyrosine-protein kinase activity. The VEGF-kinase ligand/receptor signaling system plays a key role in vascular development and regulation of vascular permeability. In case of HIV-1 infection, the interaction with extracellular viral Tat protein seems to enhance angiogenesis in Kaposi's sarcoma lesions.
Involvement in diseaseDefects in KDR are associated with susceptibility to hemangioma capillary infantile (HCI) [MIM:602089]. HCI are benign, highly proliferative lesions involving aberrant localized growth of capillary endothelium. They are the most common tumor of infancy, occurring in up to 10% of all births. Hemangiomas tend to appear shortly after birth and show rapid neonatal growth for up to 12 months characterized by endothelial hypercellularity and increased numbers of mast cells. This phase is followed by slow involution at a rate of about 10% per year and replacement by fibrofatty stroma.
Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily. Contains 7 Ig-like C2-type (immunoglobulin-like) domains. Contains 1 protein kinase domain.
Post-translational modificationsPhosphorylated. Dephosphorylated by PTPRB. Dephosphorylated by PTPRJ at Tyr-951, Tyr-996, Tyr-1054, Tyr-1059, Tyr-1175 and Tyr-1214.
Peptide Competition and Stimulation: Extracts prepared from PAE cells transfected with a chimeric receptor or from NIH3T3 cells transfected with human VEGFR 2 were stimulated with VEGF (1-4, 6) or left unstimulated (5), then were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 0.50 µg/mL ab5472 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5-6), the non phosphopeptide corresponding to the immunogen (2), a generic phospho-tyrosine containing peptide (3), or, the phosphopeptide immunogen (4). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab5472 blocks the antibody signal, and the stimulation of the phospho signal after VEGF is added (compare lanes 5 and 6), thereby demonstrating the specificity of the antibody. Note that the full-length wild type VEGFR 2 is 200 kDa (5, 6), whereas, the chimeric CSF-1-VEGFR2 is 190 kDa (1-4). The second band detected at 140 kDa in lanes 5 and 6 was observed in some but not all transfected NIH3T3 cell preparations and with multiple VEGFR 2 antibodies.
References for Anti-VEGF Receptor 2 (phospho Y1054) antibody (ab5472)
This product has been referenced in:
De Luisi A et al. Lenalidomide restrains motility and overangiogenic potential of bone marrow endothelial cells in patients with active multiple myeloma. Clin Cancer Res17:1935-46 (2011).
Read more (PubMed: 21307145) »
Moschetta M et al. Bortezomib and zoledronic acid on angiogenic and vasculogenic activities of bone marrow macrophages in patients with multiple myeloma. Eur J Cancer46:420-9 (2010).
Read more (PubMed: 19914061) »