Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)
- Datasheet
- References (16)
- Protocols
Overview
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Product nameAnti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody
See all VEGF Receptor 2 primary antibodies -
DescriptionRabbit polyclonal to VEGF Receptor 2 (phospho Y1054 + Y1059)
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Host speciesRabbit
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Tested applicationsSuitable for: ICC/IF, IHC-P, WBmore details
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Species reactivityReacts with: Mouse, Rat, Human
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Immunogen
Synthetic phospho peptide (Human)containing Tyrosines 1054 and 1059. This sequence is conserved in mouse and rat.
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Positive control
- Porcine aortic endothelial cells transfected with a chimeric receptor consisting of the extracellular portion of the CSF 1 receptor fused with the transmembrane and intracellular domains of mouse VEGFR 2.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA
BSA is IgG and protease free -
Concentration information loading... -
PurityImmunogen affinity purified
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Purification notesThe antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated VEGFR 2. The final product is generated by affinity chromatography using a VEGFR 2 derived peptide that is phosphorylated at Tyrosines 1054 and 1059.
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab5473 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| ICC/IF | 1/250. | |
| IHC-P | Use at an assay dependent concentration. PubMed: 20566746 | |
| WB | Use a concentration of 0.1 - 1 µg/ml. Detects a band of approximately 200 kDa. |
Target
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FunctionReceptor for VEGF or VEGFC. Has a tyrosine-protein kinase activity. The VEGF-kinase ligand/receptor signaling system plays a key role in vascular development and regulation of vascular permeability. In case of HIV-1 infection, the interaction with extracellular viral Tat protein seems to enhance angiogenesis in Kaposi's sarcoma lesions.
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Involvement in diseaseDefects in KDR are associated with susceptibility to hemangioma capillary infantile (HCI) [MIM:602089]. HCI are benign, highly proliferative lesions involving aberrant localized growth of capillary endothelium. They are the most common tumor of infancy, occurring in up to 10% of all births. Hemangiomas tend to appear shortly after birth and show rapid neonatal growth for up to 12 months characterized by endothelial hypercellularity and increased numbers of mast cells. This phase is followed by slow involution at a rate of about 10% per year and replacement by fibrofatty stroma.
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Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. CSF-1/PDGF receptor subfamily.
Contains 7 Ig-like C2-type (immunoglobulin-like) domains.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylated. Dephosphorylated by PTPRB. Dephosphorylated by PTPRJ at Tyr-951, Tyr-996, Tyr-1054, Tyr-1059, Tyr-1175 and Tyr-1214. -
Cellular localizationMembrane.
- Information by UniProt
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Database links
- Entrez Gene: 3791 Human
- Entrez Gene: 16542 Mouse
- Entrez Gene: 25589 Rat
- Omim: 191306 Human
- SwissProt: P35968 Human
- SwissProt: P35918 Mouse
- SwissProt: O08775 Rat
- Unigene: 479756 Human
see all -
Alternative names
- CD309 antibody
- CD309 antigen antibody
- EC 2.7.10.1 antibody
see all
Images
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Western blot - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)All lanes : Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473) at 1/100 dilution
Lane 1 : Human umbilical vein endothelial cells with Skimmed milk
Lane 2 : Human umbilical vein endothelial cells treated with 25ng/ml VEGF with Skimmed milk
Lane 3 : MDA-MB-231 with Skimmed milk
Lane 4 : MCF7 with Skimmed milk
Lysates/proteins at 30 µg per lane.
Blocking peptides at 5 % per lane.
Secondary
All lanes : HRP conjugated Goat anti-rabbit IgG at 1/5000 dilution -
Immunocytochemistry/ Immunofluorescence - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)ab5473 staining VEGF Receptor 2 (phospho Y1054 + Y1059) in 90% confluent log phase Human umbilical vein endothelial cells (panel a) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (2 ug/ml in 0.1% BSA) for 3 hours at room temperature. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/2000). Nuclei stained blue with DAPI (panel b), F-actin stained red with Alexa Fluor® 555 Rhodamine Phalloidin (panel c), merged image shown in panel d.
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Western blot - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)Peptide Competition and Stimulation: Extracts prepared from PAE cells transfected with a chimeric receptor (see Positive Controls section above for full description) were left unstimulated (1, 2) or stimulated with VEGF (3-8), then were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with a VEGFR2 pan antibody (1, 3) or 0.50µ g/mL VEGFR2 ab5473 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (2, 4, 5), the non-phosphopeptide corresponding to the immunogen (6), a generic phosphotyrosine containing peptide (7), or, the phosphopeptide immunogen (8). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar method. The data show that only the peptide corresponding to ab5473 blocks the antibody signal,
Datasheets and documents
References
This product has been referenced in:
- Jia T et al. Heteromultivalent targeting of integrin avß3 and neuropilin 1 promotes cell survival via the activation of the IGF-1/insulin receptors. Biomaterials 155:64-79 (2018). Read more (PubMed: 29169039) »
- Li X et al. Helios expression in regulatory T cells promotes immunosuppression, angiogenesis and the growth of leukemia cells in pediatric acute lymphoblastic leukemia. Leuk Res 67:60-66 (2018). Read more (PubMed: 29455107) »
