How to increase protein expression level in cell lines - (Mar/21/2006 )

Hi all,

I am a rookie in the Forum as well as the research field. Recently, I have expressed a protein in HEK cells. However, the quantity of the protein is very low (as the band only can be visualized when I concentrated the cell culture medium for 20x) that I cannot purify them out. I want to seek your expertise how to increase its expression level so that I can purify it? Thank you very much!

What vector are you currently using (complete description including ori and promoter, please)?

Is your cell line just HEK293, or is it HEK293T, HEK293E, etc.?

-Matt

I am just using pcDNA3.1(+) from Invitogen, its ori is f1 ori and the promoter is CMV.

I have used HEK293 cell lines.

By the way, is there any vector that can help to increase the expression levels?

Thank for your help!