I have mouse p50- c-terminal flag tag I PCDNA vector but I need two tags one with his as well, so it is important that his tag is also on C-terminal ad pcdna 3 only??
can I use pcdna3.1??I need to check transfection efficiency as well.
Please explain what is the thumb rule??
I cannot see exactly what it is your objective. If you use the anti flag for co-IP and your protein is a dimer you will see in the Western a band at the double of the molecular mass. Would this be enough? If you do not have any good anti-p50 Ab, i.e. if you need another flag it is almost irrelevant if you put it in the C terminal of in the N terminal, unless the homodimer interaction involves C or N terminus. In such a case use same vector for the 2 constructs. Another thumbs role would be to use a positive and a negative control in the coIP assays.
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