Human Phospho-VEGF R2/KDR/Flk-1 (Y1214) Antibody

(4 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human VEGF R2/KDR/Flk‑1 in Western blots when phosphorylated at Y1214.
  • Source
    Polyclonal Rabbit IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Phosphopeptide containing human VEGF R2/KDR/Flk-1 Y1214 site
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    2.5 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Phospho-VEGF R2/KDR/Flk‑1 (Y1214) by Western Blot. Western blot shows lysates of HUVEC human umbilical vein endothelial cells untreated (-) or treated (+) with 100 μM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-VEGF R2/KDR/Flk‑1 (Y1214) Antigen Affinity-purified Polyclonal Antibody, followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for Phospho-VEGF R2/KDR/
Flk‑1 (Y1214) at approximately 210 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
Phospho-VEGF R2/KDR/Flk-1 (Y1214) in A431 Human Cell Line. VEGF R2/KDR/Flk‑1 phosphorylated at Y1214 was detected in immersion fixed A431 human epithelial carcinoma cell line untreated (lower panel) or treated (upper panel) with pervanadate using Rabbit Anti-Human Phospho-VEGF R2/KDR/Flk‑1 (Y1214) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1766) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Intracellular Staining by Flow Cytometry
Detection of Phospho-VEGF R2/
KDR/Flk‑1 in pervanadate-treated HUVEC by Flow Cytometry.

HUVEC human umbilical vein endothelial cells were unstimulated (yellow filled histogram) or treated with 100 μM pervanadate for 15 minutes (orange filled histogram), then stained with Rabbit Anti-Human Phospho‑VEGF R2/KDR/Flk‑1
(Y1214) Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1766), or control antibody (Catalog # AB-105-C, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: VEGF R2/KDR/Flk-1

VEGF R2 (KDR/Flk-1), VEGF R1 (Flt-1) and VEGF R3 (Flt-4) belong to the class III subfamily of receptor tyrosine kinases (RTKs). All three receptors contain seven immunoglobulin-like repeats in their extracellular domains and kinase insert domains in their intracellular regions. The expression of VEGF R1, 2, and 3 is almost exclusively restricted to the endothelial cells. These receptors are likely to play essential roles in vasculogenesis and angiogenesis.

VEGF R2 cDNA encodes a 1356 amino acid (aa) residue precursor protein with a 19 aa residue signal peptide. Mature VEGF R2 is composed of a 745 aa residue extracellular domain, a 25 aa residue transmembrane domain and a 567 aa residue cytoplasmic domain. In contrast to VEGF R1 which binds both PlGF and VEGF with high affinity, VEGF R2 binds VEGF but not PlGF with high affinity. The recombinant soluble VEGF R2/Fc chimera binds VEGF with high affinity and is a potent VEGF antagonist.

  • References:
    1. Ferra, N. and R. Davis-Smyth (1997) Endocrine Reviews 18:4.
  • Long Name:
    Vascular Endothelial Growth Factor Receptor 2
  • Entrez Gene IDs:
    3791 (Human); 16542 (Mouse)
  • Alternate Names:
    CD309 antigen; CD309; EC 2.7.10; EC 2.7.10.1; Fetal liver kinase 1; fetal liver kinase-1; Flk1; Flk-1; FLK1tyrosine kinase growth factor receptor; KDR; kinase insert domain receptor (a type III receptor tyrosine kinase); Kinase insert domain receptor; Protein-tyrosine kinase receptor flk-1; soluble VEGFR2; vascular endothelial growth factor receptor 2; VEGF R2; VEGFR; VEGFR2; VEGFR-2
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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Species
Applications
Sample Type
  1. Endothelial progenitor cells transplantation attenuated blood-brain barrier damage after ischemia in diabetic mice via HIF-1?
    Authors: J Geng, L Wang, M Qu, Y Song, X Lin, Y Chen, M Mamtilahun, S Chen, Z Zhang, Y Wang, GY Yang
    Stem Cell Res Ther, 2017;8(1):163.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
  2. DSGOST inhibits tumor growth by blocking VEGF/VEGFR2-activated angiogenesis
    Oncotarget, 2016;7(16):21775-85.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Fluorescent resonance energy transfer imaging of VEGFR dimerization.
    Authors: Ahmadova Z, Yagublu V, Forg T, Hajiyeva Y, Jesenofsky R, Hafner M, Keese M
    Anticancer Res, 2014;34(5):2123-33.
    Species: Hamster
    Sample Type: Whole Cells
    Application: ICC
  4. Vascular endothelial growth factor signals through platelet-derived growth factor receptor beta in meningiomas in vitro.
    Br. J. Cancer, 2012;107(10):1702-13.
    Species: Human
    Sample Type: Cell Lysates
    Application: TaqMan Protein Assay
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