FLAG versus GST - (Jul/13/2005 )
FLAG and GST, any differences in pull-down assay?
-bigeye-
Because almost all the paper I read used GST, I am not sure whether I can use a FLAG as well. ANy body can help me? Thanks in advance
-bigeye-
Hi bigeye,
The advantage of GST is in the matrix used in purification (glutathione agarose) and the fact that it can competed off the column and purified in what is considered a gentle way.
If you use FLAG (and you can) to obtain the protein you must immunoprecipitate with an anti-FLAG antibody coupled to Protein-A or -G. Consequently, Flag is handy for detection as the anti-FLAG antibodies are very good, but if you want to purify protein than GST is definately the way to go.
Hope this helps
Scott
-Scott-