Product: Phospho-VEGFR2 (Tyr1059) Antibody
Catalog: AF3279
Description: Rabbit polyclonal antibody to Phospho-VEGFR2 (Tyr1059)
Application: WB IF/ICC
Reactivity: Human, Mouse, Rat
Prediction: Pig, Zebrafish, Bovine, Sheep, Rabbit, Dog, Xenopus
Mol.Wt.: 170kDa; 152kD(Calculated).
Uniprot: P35968
RRID: AB_2834700

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Product Info

Source:
Rabbit
Application:
WB 1:500-1:2000, IF/ICC 1:100-1:500
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
Human,Mouse,Rat
Prediction:
Pig(100%), Zebrafish(92%), Bovine(100%), Sheep(100%), Rabbit(100%), Dog(100%), Xenopus(92%)
Clonality:
Polyclonal
Specificity:
Phospho-VEGFR2 (Tyr1059) Antibody detects endogenous levels of VEGFR2 only when phosphorylated at Tyrosine 1059.
RRID:
AB_2834700
Cite Format: Affinity Biosciences Cat# AF3279, RRID:AB_2834700.
Conjugate:
Unconjugated.
Purification:
The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
Storage:
Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

CD309; CD309 antigen; EC 2.7.10.1; Fetal liver kinase 1; FLK-1; FLK1; FLK1, mouse, homolog of; Kdr; Kinase insert domain receptor (a type III receptor tyrosine kinase); Kinase insert domain receptor; KRD1; Ly73; Protein tyrosine kinase receptor FLK1; Protein-tyrosine kinase receptor flk-1; soluble VEGFR2; Tyrosine kinase growth factor receptor; Vascular endothelial growth factor receptor 2; VEGFR 2; VEGFR; VEGFR-2; VEGFR2; VGFR2_HUMAN;

Immunogens

Immunogen:
Uniprot:
Gene(ID):
Expression:
P35968 VGFR2_HUMAN:

Detected in cornea (at protein level). Widely expressed.

Description:
VEGFR-2 is a receptor tyrosine kinase of the VEGFR family. High affinity receptor for VEGF and VEGF-C. Ligand binding induces autophosphorylation and activation. Activated receptor recruits proteins including Shc, GRB2, PI3K, Nck, SHP-1 and SHP-2.
Sequence:
MQSKVLLAVALWLCVETRAASVGLPSVSLDLPRLSIQKDILTIKANTTLQITCRGQRDLDWLWPNNQSGSEQRVEVTECSDGLFCKTLTIPKVIGNDTGAYKCFYRETDLASVIYVYVQDYRSPFIASVSDQHGVVYITENKNKTVVIPCLGSISNLNVSLCARYPEKRFVPDGNRISWDSKKGFTIPSYMISYAGMVFCEAKINDESYQSIMYIVVVVGYRIYDVVLSPSHGIELSVGEKLVLNCTARTELNVGIDFNWEYPSSKHQHKKLVNRDLKTQSGSEMKKFLSTLTIDGVTRSDQGLYTCAASSGLMTKKNSTFVRVHEKPFVAFGSGMESLVEATVGERVRIPAKYLGYPPPEIKWYKNGIPLESNHTIKAGHVLTIMEVSERDTGNYTVILTNPISKEKQSHVVSLVVYVPPQIGEKSLISPVDSYQYGTTQTLTCTVYAIPPPHHIHWYWQLEEECANEPSQAVSVTNPYPCEEWRSVEDFQGGNKIEVNKNQFALIEGKNKTVSTLVIQAANVSALYKCEAVNKVGRGERVISFHVTRGPEITLQPDMQPTEQESVSLWCTADRSTFENLTWYKLGPQPLPIHVGELPTPVCKNLDTLWKLNATMFSNSTNDILIMELKNASLQDQGDYVCLAQDRKTKKRHCVVRQLTVLERVAPTITGNLENQTTSIGESIEVSCTASGNPPPQIMWFKDNETLVEDSGIVLKDGNRNLTIRRVRKEDEGLYTCQACSVLGCAKVEAFFIIEGAQEKTNLEIIILVGTAVIAMFFWLLLVIILRTVKRANGGELKTGYLSIVMDPDELPLDEHCERLPYDASKWEFPRDRLKLGKPLGRGAFGQVIEADAFGIDKTATCRTVAVKMLKEGATHSEHRALMSELKILIHIGHHLNVVNLLGACTKPGGPLMVIVEFCKFGNLSTYLRSKRNEFVPYKTKGARFRQGKDYVGAIPVDLKRRLDSITSSQSSASSGFVEEKSLSDVEEEEAPEDLYKDFLTLEHLICYSFQVAKGMEFLASRKCIHRDLAARNILLSEKNVVKICDFGLARDIYKDPDYVRKGDARLPLKWMAPETIFDRVYTIQSDVWSFGVLLWEIFSLGASPYPGVKIDEEFCRRLKEGTRMRAPDYTTPEMYQTMLDCWHGEPSQRPTFSELVEHLGNLLQANAQQDGKDYIVLPISETLSMEEDSGLSLPTSPVSCMEEEEVCDPKFHYDNTAGISQYLQNSKRKSRPVSVKTFEDIPLEEPEVKVIPDDNQTDSGMVLASEELKTLEDRTKLSPSFGGMVPSKSRESVASEGSNQTSGYQSGYHSDDTDTTVYSSEEAELLKLIEIGVQTGSTAQILQPDSGTTLSSPPV

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species
Results
Score
Pig
100
Bovine
100
Sheep
100
Dog
100
Rabbit
100
Xenopus
92
Zebrafish
92
Horse
0
Chicken
0
Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - P35968 As Substrate

Site PTM Type Enzyme
N143 N-Glycosylation
S229 Phosphorylation Q00535 (CDK5)
N245 N-Glycosylation
N318 N-Glycosylation
S711 Phosphorylation
Y801 Phosphorylation
Y822 Phosphorylation
S825 Phosphorylation
Y951 Phosphorylation P35968 (KDR)
K960 Acetylation
S982 Phosphorylation
S984 Phosphorylation
Y996 Phosphorylation P35968 (KDR)
Y1054 Phosphorylation P35968 (KDR)
K1055 Ubiquitination
Y1059 Phosphorylation P35968 (KDR)
Y1175 Phosphorylation P12931 (SRC) , P35968 (KDR)
Y1214 Phosphorylation P35968 (KDR)
T1217 Phosphorylation
Y1223 Phosphorylation
S1235 Phosphorylation
T1238 Phosphorylation
S1279 Phosphorylation
S1281 Phosphorylation
S1288 Phosphorylation
S1290 Phosphorylation
Y1305 Phosphorylation P35968 (KDR)
Y1309 Phosphorylation P35968 (KDR)
Y1319 Phosphorylation P35968 (KDR)

PTMs - P35968 As Enzyme

Substrate Site Source
P07737 (PFN1) Y129 Uniprot
P35916 (FLT4) Y1230 Uniprot
P35916 (FLT4) Y1231 Uniprot
P35916 (FLT4) Y1265 Uniprot
P35916 (FLT4) Y1333 Uniprot
P35968 (KDR) Y951 Uniprot
P35968 (KDR) Y996 Uniprot
P35968 (KDR) Y1054 Uniprot
P35968 (KDR) Y1059 Uniprot
P35968 (KDR) Y1175 Uniprot
P35968 (KDR) Y1214 Uniprot
P35968 (KDR) Y1305 Uniprot
P35968 (KDR) Y1309 Uniprot
P35968 (KDR) Y1319 Uniprot

Research Backgrounds

Function:

Tyrosine-protein kinase that acts as a cell-surface receptor for VEGFA, VEGFC and VEGFD. Plays an essential role in the regulation of angiogenesis, vascular development, vascular permeability, and embryonic hematopoiesis. Promotes proliferation, survival, migration and differentiation of endothelial cells. Promotes reorganization of the actin cytoskeleton. Isoforms lacking a transmembrane domain, such as isoform 2 and isoform 3, may function as decoy receptors for VEGFA, VEGFC and/or VEGFD. Isoform 2 plays an important 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. Modulates FLT1 and FLT4 signaling by forming heterodimers. Binding of vascular growth factors to isoform 1 leads to the activation of several signaling cascades. Activation of PLCG1 leads to the production of the cellular signaling molecules diacylglycerol and inositol 1,4,5-trisphosphate and the activation of protein kinase C. Mediates activation of MAPK1/ERK2, MAPK3/ERK1 and the MAP kinase signaling pathway, as well as of the AKT1 signaling pathway. Mediates phosphorylation of PIK3R1, the regulatory subunit of phosphatidylinositol 3-kinase, reorganization of the actin cytoskeleton and activation of PTK2/FAK1. Required for VEGFA-mediated induction of NOS2 and NOS3, leading to the production of the signaling molecule nitric oxide (NO) by endothelial cells. Phosphorylates PLCG1. Promotes phosphorylation of FYN, NCK1, NOS3, PIK3R1, PTK2/FAK1 and SRC.

PTMs:

N-glycosylated.

Ubiquitinated. Tyrosine phosphorylation of the receptor promotes its poly-ubiquitination, leading to its degradation via the proteasome or lysosomal proteases.

Autophosphorylated on tyrosine residues upon ligand binding. Autophosphorylation occurs in trans, i.e. one subunit of the dimeric receptor phosphorylates tyrosine residues on the other subunit. Phosphorylation at Tyr-951 is important for interaction with SH2D2A/TSAD and VEGFA-mediated reorganization of the actin cytoskeleton. Phosphorylation at Tyr-1175 is important for interaction with PLCG1 and SHB. Phosphorylation at Tyr-1214 is important for interaction with NCK1 and FYN. Dephosphorylated by PTPRB. Dephosphorylated by PTPRJ at Tyr-951, Tyr-996, Tyr-1054, Tyr-1059, Tyr-1175 and Tyr-1214.

The inhibitory disulfide bond between Cys-1024 and Cys-1045 may serve as a specific molecular switch for H(2)S-induced modification that regulates KDR/VEGFR2 function.

Subcellular Location:

Cell junction. Endoplasmic reticulum.
Note: Localized with RAP1A at cell-cell junctions (By similarity). Colocalizes with ERN1 and XBP1 in the endoplasmic reticulum in endothelial cells in a vascular endothelial growth factor (VEGF)-dependent manner (PubMed:23529610).

Cell membrane>Single-pass type I membrane protein. Cytoplasm. Nucleus. Cytoplasmic vesicle. Early endosome.
Note: Detected on caveolae-enriched lipid rafts at the cell surface. Is recycled from the plasma membrane to endosomes and back again. Phosphorylation triggered by VEGFA binding promotes internalization and subsequent degradation. VEGFA binding triggers internalization and translocation to the nucleus.

Secreted.

Secreted.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionSubcellular location
Tissue Specificity:

Detected in cornea (at protein level). Widely expressed.

Subunit Structure:

Homodimer in the presence of bound dimeric VEGFA, VEGFC or VEGFD ligands; monomeric in the absence of bound ligands. Can also form heterodimers with FLT1/VEGFR1 and KDR/VEGFR2. Interacts (tyrosine phosphorylated) with LFYN, NCK1, PLCG1. Interacts (tyrosine-phosphorylated active form preferentially) with DAB2IP (via C2 domain and active form preferentially); the interaction occurs at the late phase of VEGFA response and inhibits KDR/VEGFR2 activity. Interacts with SHBSH2D2A/TSAD, GRB2, MYOF, CBL and PDCD6. Interacts (via C-terminus domain) with ERN1 (via kinase domain); the interaction is facilitated in a XBP1 isoform 1- and vascular endothelial growth factor (VEGF)-dependent manner in endothelial cells. Interacts (via juxtamembrane region) with chaperone PDCL3 (via thioredoxin fold region); the interaction leads to increased KDR/VEGFR2 abundance through inhibition of its ubiquitination and degradation. Interacts (tyrosine phosphorylated) with CCDC88A/GIV (via SH2-like region); binding requires autophosphorylation of the KDR/VEGFR2 C-terminal region.

(Microbial infection) Interacts with HIV-1 Tat.

Family&Domains:

The second and third Ig-like C2-type (immunoglobulin-like) domains are sufficient for VEGFC binding.

Belongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.

Research Fields

· Cellular Processes > Cellular community - eukaryotes > Focal adhesion.   (View pathway)

· Environmental Information Processing > Signal transduction > MAPK signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Ras signaling pathway.   (View pathway)

· Environmental Information Processing > Signal transduction > Rap1 signaling pathway.   (View pathway)

· Environmental Information Processing > Signaling molecules and interaction > Cytokine-cytokine receptor interaction.   (View pathway)

· Environmental Information Processing > Signal transduction > PI3K-Akt signaling pathway.   (View pathway)

· Human Diseases > Drug resistance: Antineoplastic > EGFR tyrosine kinase inhibitor resistance.

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

References

1). Effect of Extracellular Ribonucleic Acids on Neurovascularization in Osteoarthritis. Advanced Science, 2023 (PubMed: 37395388) [IF=15.1]

Application: WB    Species: Mouse    Sample: TG cells

Figure 8 The exRNA‐VEGF complex promotes angiogenesis and neurogenesis through action on VEGFR2. A) Representative microscopy images of TG cells showing the dendritic length and interactions after 24 h treatment. B) Representative light microscopy images of EPCs showing migrated cells, cell migration, and tube formation after 24 h treatment. C,D) Quantification of dendritic length and interactions of TGs. E,F) Quantification of the numbers and distance of migrated EPCs. G) Western blot results showing expression of VEGFR2 and p‐VEGFR2 of TG cells and EPCs after treatment for 24 h. Scale bars = 50 µm (A), 100 µm (top and bottom in B), and 500 µm (middle in B). Data are shown as the means and standard deviations; p < 0.05 (n = 3). Different letters indicate statistically significant differences.

2). β-Sitosterol Inhibits Rheumatoid Synovial Angiogenesis Through Suppressing VEGF Signaling Pathway. Frontiers in Pharmacology, 2022 (PubMed: 35295740) [IF=5.6]

Application: WB    Species: Human    Sample: HUVECs

FIGURE 4 The effect of VEGF signaling pathway in TNF-α-induced HUVECs regulated by BSS. (A) GSEA analysis of KEGG signaling pathway in the GSE121894 dataset. The structural interactions between VEGFR2 and BSS (B) and between VEGFR2 and axitinib (C) were described by molecular simulation, and binding affinity of VEGFR2 with BSS and axitinib is shown in Table 1. (D) p-VEGFR2 and VEGFR2 protein levels in HUVECs (on TNF-α 20 ng/ml for 1 h) intervened with BSS dosage groups were detected by immunoblotting and the relative expression levels (E) of proteins were corrected by GAPDH (n = 3). (F) p-VEGFR2 and VEGFR2 protein levels in HUVECs intervened with BSS and axitinib were detected by immunoblotting, and the relative expression levels (G) of proteins were corrected by GAPDH (n = 3). All data are shown as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, n.s. = not significant.

Application: IHC    Species: Mice    Sample: synovial tissues

FIGURE 6 Effect of BSS on immunohistochemistry of CD31, VEGFR2, and P-VEGFR2 on experimental arthritis. (A) CD31 immunohistochemical staining of the ankle joints in each group, and the optical density value (B) analysis using ImageJ (n = 6) (×200, scale bar = 50 μm). (C) VEGFR2 immunohistochemical staining of the ankle joints in each group, and the optical density value (D) analysis using ImageJ (n = 6) (×200, scale bar = 50 μm). (E) p-VEGFR2 immunohistochemical staining of the ankle joints in each group, and (F) the optical density value analysis using ImageJ (n = 6) (×200, scale bar = 50 μm). All data are shown as the mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, n.s. = not significant.

3). SU5416 attenuated lipopolysaccharide-induced acute lung injury in mice by modulating properties of vascular endothelial cells. Drug Design Development and Therapy, 2019 (PubMed: 31213766) [IF=4.8]

Application: WB    Species: mouse    Sample: lung

Figure 3 |TLR4/NF-κB signaling was involved in the progression of LPS-stimulated ALI. (A) Immunohistochemical staining of CD31 in LPS-stimulated WT and TLR4−/- mice after treatment with DXM and/or SU5416+DXM.. (B) Mice were treated with DXM and/or SU5416 for 12 hours, and the expressions of TLR4, p-p65, NF-κB, p-VEGFR2,VEGF2R, p53, Bcl-2, and β-actin in lung tissues were detected with Western blot

4). The role of Apatinib combined with Paclitaxel (aluminum binding type) in drug-resistant ovarian cancer. Journal of Ovarian Research, 2020 (PubMed: 32958014) [IF=4.0]

Application: WB    Species: human    Sample: SKOV-3cell

Fig. 1 |Effect of nab-P and AP on SKOV-3/DDP. a the nab-P and AP’s half inhibition concentration to SKOV-3/DDP (24, 48 and 72 h), b the Bax, bcl-2, pVEGFR-2 and MMP-2 protein expression of different groups by Western blot (WB)

Application: IF/ICC    Species: human    Sample: SKOV-3cell

Fig. 1 |Effect of nab-P and AP on SKOV-3/DDP. a the nab-P and AP’s half inhibition concentration to SKOV-3/DDP (24, 48 and 72 h), b the Bax, bcl-2, pVEGFR-2 and MMP-2 protein expression of different groups by Western blot (WB),c the Bax, bcl-2, p-VEGFR-2 and MMP-2 protein expression of different groups by Immunofluorescence (IF)

5). Endostar Synergizes with Radiotherapy to Inhibit Angiogenesis of Cervical Cancer in a Subcutaneous Xenograft Mouse Model. Frontiers in Bioscience - Elite, 2022 (PubMed: 36042171)

Application: WB    Species: Human    Sample: HUVECs

Fig. 7. The effect of Endostar on HIF-1α/VEGFR2/PI3K/AKT/DNA-PKcs pathway activation after radiotherapy in HUVECs. Western blotting analysis revealed that in comparison with RT monotherapy, Endostar+RT significantly inhibited the phosphorylation of VEGFR, PI3K, AKT, and DNA-PKcs in HUVECs (p < 0.05). Simultaneously, H2AX expression decreased significantly (p < 0.05).

6). Apatinib Induces Ferroptosis of Glioma Cells through Modulation of the VEGFR2/Nrf2 Pathway. Oxidative Medicine and Cellular Longevity, 2022 (PubMed: 35602105)

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