Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)
Key features and details
- Rabbit polyclonal to VEGF Receptor 2 (phospho Y1054 + Y1059)
- Suitable for: ICC/IF, WB
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody
See all VEGF Receptor 2 primary antibodies -
Description
Rabbit polyclonal to VEGF Receptor 2 (phospho Y1054 + Y1059) -
Host species
Rabbit -
Tested applications
Suitable for: ICC/IF, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat
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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
- IF: VEGF treated HUVEC whole cells. WB: VEGF treated HUVEC, MDA-MB-231 and MCF7 whole cell extract.
Properties
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Form
Liquid -
Storage instructions
Shipped 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. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.1% BSA, 50% Glycerol -
Concentration information loading... -
Purity
Immunogen affinity purified -
Purification notes
The 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. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Images
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Immunocytochemistry/ Immunofluorescence - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473)
Immunofluorescence analysis of HUVEC cells labelling VEGF Receptor 2 (phospho Y1054 + Y1059) (PAnel a: green) using ab5473 at 2 ug/ml in 0.1% BSA for 3 hours at room temperature, follwed by Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal at 1/2000 dilution. Pbel b: Nuclei stained with DAPI (blue), Panel c: F-actin stained with Alexa Fluor® 555 Rhodamine Phalloidin (red), Panel d: Merged images. The images were captured at 60X magnification.
Prior antibody incubation, HUVEC (Human umbilical vein endothelial cell line) cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes and blocked with 1% BSA for 1 hour at room temperature, followed by treatment with 100 ng of VEGF for 30 minutes. Assay was done on 90% confluent log phase HUVEC cells.
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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 1/1000 dilution
Lane 1 : HUVEC (Human umbilical vein endothelial cell line) whole cell extract with Skimmed milk
Lane 2 : HUVEC cells treated with 25ng/ml VEGF for 5 minutes with Skimmed milk
Lane 3 : MDA-MB-231 (human breast adenocarcinoma cell line) whole cell extract with Skimmed milk
Lane 4 : MCF7 (human breast adenocarcinoma cell line) whole cell extract with Skimmed milk
Lysates/proteins at 30 µg per lane.
Blocking peptides at 5 % per lane.
Secondary
All lanes : Goat anti-rabbit IgG HRP conjugate at 1/5000 dilutionA 130 kDa band corresponding to VEGFR (pYpY1054/1059) was observed across cell lines tested.
Protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel, XCell SureLock™ Electrophoresis System and Novex® Sharp Pre-Stained Protein Standard.
Resolved proteins were then transferred onto a nitrocellulose membrane by over night wet transfer.
Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate.
