could FLAG-tagged protein be used in the Luc assay? (View forum version)



gyma

Posted 16 November 2009 - 05:11 AM

Hi, everyone. I have several expression plasmids which are all flag-tagged and I have already used them in my Luc assay. I am wondering if FLAG affects the result? Is there anyone else ever using a tagged plasmid in the Luc assay? Thanks in advance.

laurequillo

Posted 07 December 2009 - 09:57 PM

Hi there,

they should not affect your resulst at all. I do it eveyday.

But just to be sure it would be good to check some untagged vector or some vectors with a different tag. The reason is not because flag-tag could affect the luc assay, the reason is because sometimes different tags or different location of the tag (N vs C terminal tags) can affect the activity of the protein. So, if you have any doubt, it would be good to check the same protein with different tags, or even better the untagged protein to see if they have the same behaviour.