2 replies to this topic

[Biochem_newbie]

Hi everyone,

I'm currently using 293T cells to over-express my protein of interest.
I know 293T cells are used because they are easily transfectable.

My concern is that it feels like everytime I transfect, and run a Western, the expression patterns of the bands are completely different, even with same amount of DNA and protein loaded.

I am wondering, does the variation in confluency of the cells play a role in the differing protein expression patterns each time I run a Western?

Would really appreciate your advice!

Thank you

[bob1]

Yes it can.  Depends on the protein though.

[ascacioc]

This article might help you understand the differences you see:
http://www.ncbi.nlm....pubmed/21354312