Anti-p-VEGFR2 (Tyr1059) Antibody, clone 4C22-H1 ZooMAb^® Rabbit Monoclonal

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ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

Clone 4C22-H1 is a ZooMAb^® rabbit recombinant monoclonal antibody that specifically detects VEGFR2 phosphorylated on tyrosine 1059.

KLH-conjugated linear peptide corresponding to 12 amino acids surrounding phosphotyrosine 1059 from the C-terminal half of human VEGFR2.

Quality Control Testing

Evaluated by Western Blotting in lysate from porcine aortic endothelial cells expressing chimeric VEGFR2 and stimulated with CSF-1.

Western Blotting Analysis: A 1:10,000 dilution of this antibody detected phospho-VEGFR2 (Tyr1059) in lysate from porcine aortic endothelial cells expressing chimeric VEGFR2 and stimulated with CSF-1 (40 ng/ml for 15 min).

Tested Applications

Peptide Inhibition Assay: Target band detection in lysate from porcine aortic endothelial cells expressing chimeric VEGFR2 and stimulated with CSF-1 (40 ng/ml for 15 min)., and in Human umbilical vein endothelial cells (HUVEC) treated with VEGF were prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected VEGF Receptor phosphorylated on tyrosine 1059 in overnight serum starved porcine aortic endothelial cells expressing chimeric VEGFR2 and stimulated with CSF-1.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Vascular endothelial growth factor receptor 2 (UniProt: P35968; also known as EC:2.7.10.1, VEGFR-2, Fetal liver kinase 1, FLK-1, Kinase insert domain receptor, KDR, Protein-tyrosine kinase receptor flk-1, CD309) is encoded by the KDR (also known as FLK1, VEGFR2) gene (Gene ID: 3791) in human. VEGFR-2 is highly glycosylated protein that is synthesized with a signal peptide (aa 1-19), which is subsequently cleaved off to produce the mature form that contains an extracellular domain (aa 20-764), a transmembrane domain (aa 765-785), and a cytoplasmic domain (aa 786-1356). In the absence of a bound ligand, it exists as a monomer. However, it homodimerizes upon binding to ligands and undergoes autophosphorylation and activation. Tyrosine residues 951, 996, 1054, and 1059 have been identified as autophosphorylation sites. Tyrosine residues 1059 and 951 are reported to be essential for VEGF-induced endothelial cell proliferation and migration, respectively. Y1059F mutation results in the cOmplete^™ loss of VEGFR2 -mediated intracellular Ca2+ mobilization and MAPK phosphorylation. VEGFR2 acts as a cell-surface receptor for VEGFA, VEGFC, and VEGFD and has the protein tyrosine kinase activity. It plays an essential role in the regulation of angiogenesis, vascular development, vascular permeability, and embryonic hematopoiesis. It promotes proliferation, survival, migration, and differentiation of endothelial cells.  Three isoforms of VEGFR-2 have been described that are produced by alternative splicing. Binding of VEGF to isoform 1 LEADS^™ to the activation of several signaling cascades, including the activation of ERK1, ERK2, and AKT1. It is also required for VEGFA-mediated induction of NOS2 and NOS3, leading to the production of the nitric oxide by endothelial cells. Isoforms 2 and 3 lack a transmembrane domain and may function as decoy receptors for VEGFA, VEGFC, and VEGFD. Isoform 2 is reported to play a 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. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, Affinity^™, reproducibility, and stability over conventional monoclonals. (Ref.: Wang, X., et al. (2020). Front. Dev. Biol. 8; :599281; Zheng, H., et al. (2001). J. Biol. Chem. 276(35); 32714-32719; Rahimi, N., et al. (2000). J. Biol. Chem. 275(22); 16986-16992; Dougher, M., and Terman, BI. (1999). Oncogene. 18(8); 1619-1627; Kroll, J., and Waltenberger, J. (1998). Biochem. Biophys. Res. Commun. 252(3); 743-746).

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

Reconstitute lyophilized antibody pellet with 25μL of ultrapure water or Phosphate Buffered Saline (PBS). Please refer to our reconstitution protocol and the specific application guidance on the suggested starting dilutions and sample type.

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Affinity is a trademark of Mine Safety Appliances Co.

LEADS is a trademark of Compugen

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

cOmplete is a trademark of Roche

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.