Anti-Phospho-VEGF Receptor 2-Y1175 Rabbit Polyclonal Antibody for WB - P35968

Core Product Specifications and Parameters

Biological Background: VEGF Receptor 2 Function and Localization

• VEGF Receptor 2 (VEGFR-2), also known as KDR, FLK-1, or CD309, is a receptor tyrosine kinase that serves as the primary signaling receptor for VEGFA, VEGFC, and VEGFD. It is a member of the immunoglobulin superfamily and contains multiple extracellular immunoglobulin-like domains.
• VEGFR-2 plays a critical role in angiogenesis and vascular development, promoting endothelial cell proliferation, survival, migration, and differentiation. It is essential for embryonic hematopoiesis and vascular permeability.
• Upon ligand binding, VEGFR-2 activates downstream signaling cascades, including the MAPK/ERK and AKT1 pathways, through phosphorylation of key tyrosine residues such as Y1175. Phosphorylation at Y1175 leads to activation of PLCG1 and subsequent production of second messengers like DAG and IP3.
• Isoforms lacking the transmembrane domain (isoform 2 and 3) may function as decoy receptors, negatively regulating lymphangiogenesis by sequestering VEGFA and VEGFC and preventing their binding to FLT4. VEGFR-2 also modulates signaling by forming heterodimers with FLT1 and FLT4.
• Subcellularly, VEGFR-2 is located at the cell membrane, cell junctions, endoplasmic reticulum, cytoplasm, nucleus, and early endosomes. It can also be secreted. This diverse localization reflects its dynamic trafficking and signaling roles.
• Post-translational modifications include phosphorylation (as targeted by this antibody), glycosylation, and ubiquitination. These regulate receptor activation, trafficking, and turnover.
• VEGFR-2 is widely expressed, with detection in the cornea and throughout many tissues, underscoring its broad role in vascular biology.

Experimental Guidance and Technical Tips

• The immunogen, a synthetic phosphorylated peptide around Y1175, ensures specificity for the phosphorylated form of VEGFR-2. For Western blot, consider stimulating cells with VEGFA to enhance phosphorylation signal and validate with appropriate negative controls (untreated or phosphatase-treated lysates).
• This antibody can be used in ELISA; consider optimizing coating concentration and pairing with a detection antibody for a sandwich assay targeting phospho-VEGFR-2.
• Since cross-reactivity is reported for human, mouse, and rat, validate the antibody in the relevant species and tissue systems. Include known positive controls, such as VEGF-treated HUVECs for human, or equivalent primary endothelial cells for mouse/rat.

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