Review of Phospho-EGFR antibody from Cell Signaling Technology

Biochemistry and Cancer Biology
University of Toledo Health Science Campus
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

Phospho-EGF Receptor (Tyr1068) (D7A5) XP® Rabbit mAb

Catalog Number:

3777

This antibody was used in a study to examine phosphorylation and signaling of EGF Receptor. U251 human glioma cells were serum starved overnight, incubated in 10ug/ml Cycloheximide to block new protein synthesis and then treated with EGF ligand to stimulate receptor internalization and phosphorylation.

Experimental Design and Results Summary

Applications

Western Blot

Sample

U251 human glioma cell lysate

Primary Incubation

1:1000

Blocking Agent

5% Milk in TBS- 0.2% Tween

Secondary Incubation

Goat anti-rabbit antibody; 1/3000

Tertiary Incubation

None

Detection

Femto HRP Chemiluminescent substrate Kit

Results Summary

Antibody works very well! Antibody identifies a specific band. Result shows phosphorylation of EGF Receptor by 30 minutes of EGF ligand stimulation and then a gradual decline in receptor phosphorylation. This is consistent with what is known about the EGF Receptor phosphorylation events in most cell types

Additional Notes

None

Related Categories

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Summary

The Good

Antibody works well

The Bad

None

The Bottom Line

Antibody works very well

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