I've been following this forum for a while (and it has always proved useful) but this is my first post on the site.
I've been having numerous problems cloning and expressing a gene of interest in a pCMV-Tag3a mammalian expression vector for quite some time now.
I've had my construct sequenced and the N-terminal myc tag is in frame with my protein of interest.
However, upon transient transfection into HEK-293Ts, I am unable to detect expression the construct through Western blotting. I use a pCMV-Tag3a mammalian expression vector containing another protein (generated by someone else in the lab a number of years ago) as a positive control and can detect this through Western blotting with no problems. So I do not feel I have an obvious transfection issue.
I'm not quite sure where to go from here...
Appreciate any advice or help.
IXAMember Since 24 Sep 2012
Offline Last Active Mar 07 2013 12:30 PM
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