Alternatives to MYC epitope tag

Seth M Fisher Seth_M_Fisher%notes at sb.com
Thu Nov 17 10:03:45 EST 1994


As an alternative to myc epitope the following was suggested

>I can suggest you the FLAG epitope, which can be cleaved off by
>enterokinase. Two monoclonal antibodies against FLAG are available from
>IBI/Kodak. Antibody M1 recognizes the epitope when it is exposed at the
>N-terminal of a protein. Antibody M2 recognizes it when it is internal or
>C-terminal. FLAG is 8 amino acids long.

My experience with the FLAG epitope for purification from E. Coli has not been 
good.
 The binding capacity of the M2 resin was less than 100 ug/ml. This would have
been OK if the antibody did not also recognize several other proteins, most 
notably
chaperonin.
Also, I could not cleave the FLAG off with enterokinase. The antibody did work
great for Westerns.
We switched to a hexa-his tag with more favorable results.
I would be interested in hearing in a little more detail of some success 
stories with the
FLAG technology.




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