Human Phospho-VEGFR2/KDR/Flk-1 (Y1214) Antibody
R&D Systems, part of Bio-Techne | Catalog # AF1766
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human, Hamster
Applications
Validated:
CyTOF-ready, Immunocytochemistry, Intracellular Staining by Flow Cytometry, Western Blot
Cited:
ICC, TaqMan Protein Assay, Western Blot
Label
Unconjugated
Antibody Source
Polyclonal Rabbit IgG
Product Specifications
Immunogen
Phosphopeptide containing human VEGFR2/KDR/Flk-1 Y1214 site
Specificity
Detects human VEGFR2/KDR/Flk‑1 in Western blots when phosphorylated at Y1214.
Clonality
Polyclonal
Host
Rabbit
Isotype
IgG
Scientific Data Images for Human Phospho-VEGFR2/KDR/Flk-1 (Y1214) Antibody
Detection of Human Phospho-VEGFR2/KDR/Flk‑1 (Y1214) by Western Blot.
Western Blot shows lysates of A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 μM pervanadate (PV) for 10 minutes. PVDF membrane was probed with 1 µg/ml of Rabbit Anti-Human Phospho-VEGFR2/KDR/Flk-1 (Y1214) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1766) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for VEGFR2/KDR/Flk-1 at approximately 230 kDa (as indicated). This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.
Phospho-VEGFR2/KDR/Flk-1 (Y1214) in A431 Human Cell Line.
VEGFR2/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-VEGFR2/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; NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of Phospho-VEGFR2/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-VEGFR2/KDR/Flk-1 (Y1214) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1766), or control antibody (AB-105-C, open histogram), followed by Phycoerythrin-conjugated Anti-Rabbit IgG Secondary Antibody (F0110). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.Applications for Human Phospho-VEGFR2/KDR/Flk-1 (Y1214) Antibody
Application
Recommended Usage
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
Sample: Immersion fixed A431 human epithelial carcinoma cell line treated with pervanadate
Sample: Immersion fixed A431 human epithelial carcinoma cell line treated with pervanadate
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
Sample: HUVEC human umbilical vein endothelial cells treated with pervanadate, fixed with paraformaldehyde, and permeabilized with methanol
Sample: HUVEC human umbilical vein endothelial cells treated with pervanadate, fixed with paraformaldehyde, and permeabilized with methanol
Western Blot
1 µg/mL
Sample: A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 μM pervanadate (PV) for 10 minutes
Sample: A431 human epithelial carcinoma cell line untreated (-) or treated (+) with 100 μM pervanadate (PV) for 10 minutes
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
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: VEGFR2/KDR/Flk-1
VEGFR2 (KDR/Flk-1), VEGFR1 (Flt-1) and VEGFR3 (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 VEGFR1, 2, and 3 is almost exclusively restricted to the endothelial cells. These receptors are likely to play essential roles in vasculogenesis and angiogenesis.
VEGFR2 cDNA encodes a 1356 amino acid (aa) residue precursor protein with a 19 aa residue signal peptide. Mature VEGFR2 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 VEGFR1 which binds both PlGF and VEGF with high affinity, VEGFR2 binds VEGF but not PlGF with high affinity. The recombinant soluble VEGFR2/Fc chimera binds VEGF with high affinity and is a potent VEGF antagonist.
References
- Ferra, N. and R. Davis-Smyth (1997) Endocrine Reviews 18:4.
Long Name
Vascular Endothelial Growth Factor Receptor 2
Alternate Names
CD309, Flk-1, Flk1, KDR, KRD1, Ly73, VEGF R2
Gene Symbol
KDR
Additional VEGFR2/KDR/Flk-1 Products
Product Documents for Human Phospho-VEGFR2/KDR/Flk-1 (Y1214) Antibody
Product Specific Notices for Human Phospho-VEGFR2/KDR/Flk-1 (Y1214) Antibody
For research use only
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