Good Antibody to Detect Activated VEGFR2

September 18, 2018

The Cell and Molecular Biology
The Scripps Research Institute
Assistant Professor

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Company:

Cell Signaling

Product Name:

Phospho-VEGF Receptor 2 (Tyr951) Antibody

Catalog Number:

2471

VEGFR2 is the main receptor mediating functions of VEGF (vascular endothelial growth factor). Binding of VEGF to VEGFR2 induces cell signaling, which begins with the autophosphorylation of VEGFR2 and its functional activation. One of VEGFR2 phosphorylation sites is located at Tyr951, which is reflected in the name of the rabbit antibody #2471 from Cell Signaling. We used this antibody to analyze activation status of VEGFR2 in the angiogenic tumor vasculature in human tumor xenografts grown in immunodeficient mice. By using murine endothelial cells treated with recombinant VEGF, we demonstrated that the antibody 2471 nicely recognized activated VEGFR2 (pVEGFR2).

Experimental Design and Results Summary

Applications

Western Blot

Sample

Lysates of murine endothelial cells bEND.3, loaded at 20 micrograms per lane

Primary Incubation

Primary antibody at a final dilution 1:1000; overnight incubation in blocking buffer; cold room; gentle rocking

Blocking Agent

5% BSA in PBS-0.05% Tween; 45-60 min, RT, gentle rocking

Secondary Incubation

Goat anti rabbit IgG conjugated with HRP; 1:3,000; 1 hr; RT; gentle rocking

Tertiary Incubation

N/A

Detection

West Pico

Results Summary

In our hands, the rabbit antibody #2471 nicely recognized phosphorylated VEGFR2 in the murine endothelial cells treated for 5 min with VEGF at 50 or 200 ng/mL. The recognized protein band has an expected mol. wt. of ~150 kDa.

DOI or PMID #

N/A

Additional Notes

The image demonstrates that the antibody detected a protein band when the endothelial cells were treated with VEGF. The band has an apparent mol. wt. ~150 kDa, consistent with the expected mol. wt. of VEGFR2 (~150-180 kDa).

Image Gallery

Summary

The Good

The antibody appears sensitive and capable of recognizing phosphorylated VEGFR2 in relatively small amounts of cell lysate.

The Bad

Along with pVEGFR2, the antibody 2471 produces a nimber of additional non-specific bands.

The Bottom Line

The rabbit antibody #2471 from Cell Signaling is suitable for detection in vitro of activated VEGFR2 in the endothelial cells by western blotting (under reducing conditions).

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