pAKT Western Blot Detection

June 07, 2022

BlueskyReddit
NanoEngineering
UC San Diego NanoEngineering Department
Graduate Student

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Company:

Cell Signaling

Product Name:

Phospho-Akt (Ser473) (D9E) XP®

Catalog Number:

4060

Clone Number:

N/A

I used this antibody on epithelial cells starved for 4 hours (0%FBS) and treated with eIF4E siRNA for 24 hours with F-12k medium (2% FBS). My goal was to activate AKT pathway and see the expression of eIF4E. I got great results and band was detected at 30kDa. I incubated the membrane overnight at 4 °C and the secondary antibody was used for 2 hours at RT.

Experimental Design and Results Summary

Applications

Western Blot

Sample

Epithelial cells

Primary Incubation

1:1000 overnight at 4 °C

Blocking Agent

BSA 5% in 0.1% TBST

Secondary Incubation

Rabbit RT for 2 hours

Tertiary Incubation

Didn't use.

Detection

ChemiDOC

Results Summary

I used this antibody on epithelial cells starved for 4 hours (0%FBS) and treated with eIF4E siRNA for 24 hours with F-12k medium (2% FBS). My goal was to activate AKT pathway and see the expression of eIF4E. I got great results and band was detected at 30kDa. I incubated the membrane overnight at 4 °C and the secondary antibody was used for 2 hours at RT.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Reliable and good signal.

The Bad

No bad things.

The Bottom Line

I really recommend it.

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