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The Tandem Affinity Purification (TAP) Method: A General Procedure of Protein Complex Purification

2001; Elsevier BV; Volume: 24; Issue: 3 Linguagem: Inglês

10.1006/meth.2001.1183

1095-9130

Óscar Puig, Friederike Caspary, Guillaume Rigaut, Berthold Rutz, Emmanuelle Bouveret, Elisabeth Bragado‐Nilsson, Matthias Wilm, Bertrand Séraphin,

Ubiquitin and proteasome pathways

Identification of components present in biological complexes requires their purification to near homogeneity. Methods of purification vary from protein to protein, making it impossible to design a general purification strategy valid for all cases. We have developed the tandem affinity purification (TAP) method as a tool that allows rapid purification under native conditions of complexes, even when expressed at their natural level. Prior knowledge of complex composition or function is not required. The TAP method requires fusion of the TAP tag, either N- or C-terminally, to the target protein of interest. Starting from a relatively small number of cells, active macromolecular complexes can be isolated and used for multiple applications. Variations of the method to specifically purify complexes containing two given components or to subtract undesired complexes can easily be implemented. The TAP method was initially developed in yeast but can be successfully adapted to various organisms. Its simplicity, high yield, and wide applicability make the TAP method a very useful procedure for protein purification and proteome exploration.