How can I purify a TAP-tagged yeast protein?
A description of the purification of the TAP-tagged proteins expressed by the TAP-tagged yeast strains can be found at: http://www-db.embl-heidelberg.de/jss/servlet/de.embl.bk.wwwTools.GroupLeftEMBL/ExternalInfo/seraphin/TAPpurification.html
A brief overview of the process is:
1. Proteins are expressed in the yeast under control of the native promoters.
2. Run on IgG column which binds protein A
3. Cleave with TEV protease which removes the tagged protein from the column leaving the ORF and calmodulin binding protein.
4. Run on column that binds calmodulin
5. End product is pure protein that you wish to analyze.