Excellent Antibody To Detect Phosphorylated EGFR In Human Tumor Cells

The Cell and Molecular Biology
The Scripps Research Institute
Assistant Professor

Overall

Quality of Results

Ease-of-Optimization

What do these ratings mean?
Write a Review

Company:

Cell Signaling

Product Name:

Phospho-EGF Receptor (Tyr1068) Rabbit Antibody

Catalog Number:

2234

Functional activity of epidermal growth factor receptor (EGFR) is associated with the autophosphorylation of several tyrosine (Y) residues in the C-terminal domain. Detection of phosphorylated EGFR is important in cell biology, especially in studying cancer cells. We used Cell Signaling rabbit antibody 2234 that detects phosphorylation at Y1068 to determine the levels of EGFR activation and the efficiency of EGFR inhibitors in preventing EGFR phosphorylation.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Lysates of human tumor cells; 20 microgram per lane

Primary Incubation

Antibody at a final concentration of 1 microgram/ml, ON incubation with gentle rocking in a cold room

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

Rabbit antibody 2234 produces a single protein band of approximately 180 kDa, which corresponds to expected mol. mass of EGFR.

DOI or PMID #

N/A

Additional Notes

The image demonstrates that in the presence of an EGFR inhibitor, Erlotinib, detection of phosphorylated EGFR (pEGFR) is significantly reduced, indicating that Erlotinib efficiently inhibited EGFR activation. Similar levels of total EGFR in the same samples confirm equal loading of EGFR protein.

Image Gallery

Summary

The Good

The antibody appears highly specific and sensitive.

The Bad

None detected.

The Bottom Line

The rabbit antibody 2234 from Cell Signaling is highly recommended for detection by western blotting of phosphorylated EGFR in lysates of human tumor cells.

Share your experience with other scientists. Write a Review! »

Join the discussion