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Cell confluency, transfection, protein expression for 293T cells

Transfection Protein expression 293T cells Cell confluency

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#1 Biochem_newbie

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Posted 15 September 2012 - 12:10 PM

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

#2 bob1

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Posted 15 September 2012 - 01:29 PM

Yes it can. Depends on the protein though.

#3 ascacioc

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Posted 19 September 2012 - 12:48 PM

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





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