Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody

• WB

Supplier Data

Western blot - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (AB5473)

A 150 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.

All lanes:

Western blot - Anti-VEGF Receptor 2 (phospho Y1054 + Y1059) antibody (ab5473) at 1/1000 dilution

Lanes 1 - 4:

Goat anti-rabbit IgG HRP conjugate at 1/5000 dilution

Lane 1:

HUVEC (Human umbilical vein endothelial cell line) whole cell extract at 30 µg

Lane 2:

HUVEC cells treated with 25ng/ml VEGF for 5 minutes at 30 µg

Lane 3:

MDA-MB-231 (human breast adenocarcinoma cell line) whole cell extract at 30 µg

Lane 4:

MCF7 (human breast adenocarcinoma cell line) whole cell extract at 30 µg

Predicted band size: 151 kDa

false

• ICC

Supplier Data

Immunocytochemistry - 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.