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

Methods ◽  
2001 ◽  
Vol 24 (3) ◽  
pp. 218-229 ◽  
Author(s):  
Oscar Puig ◽  
Friederike Caspary ◽  
Guillaume Rigaut ◽  
Berthold Rutz ◽  
Emmanuelle Bouveret ◽  
...  
Keyword(s):  
Protein Complex ◽  
Affinity Purification ◽  
General Procedure ◽  
Tandem Affinity Purification

scholarly journals An alternative tandem affinity purification strategy applied to Arabidopsis protein complex isolation

2005 ◽  
Vol 41 (5) ◽  
pp. 767-778 ◽  
Author(s):  
Vicente Rubio ◽  
Yunping Shen ◽  
Yusuke Saijo ◽  
Yule Liu ◽  
Giuliana Gusmaroli ◽  
...  
Keyword(s):  
Protein Complex ◽  
Affinity Purification ◽  
Tandem Affinity Purification ◽  
Arabidopsis Protein

Tandem affinity purification of theCandida albicans septin protein complex

Yeast ◽  
2004 ◽  
Vol 21 (12) ◽  
pp. 1025-1033 ◽  
Author(s):  
Aki Kaneko ◽  
Takashi Umeyama ◽  
Nozomu Hanaoka ◽  
Brian C. Monk ◽  
Yoshimasa Uehara ◽  
...  
Keyword(s):  
Protein Complex ◽  
Affinity Purification ◽  
Tandem Affinity Purification

Interaction proteomics: characterization of protein complexes using tandem affinity purification–mass spectrometry

2010 ◽  
Vol 38 (4) ◽  
pp. 883-887 ◽  
Author(s):  
Pamela Völkel ◽  
Perrine Le Faou ◽  
Pierre-Olivier Angrand
Keyword(s):  
Protein Complex ◽  
Mammalian Cells ◽  
Protein Identification ◽  
Biochemical Characterization ◽  
Protein Complexes ◽  
Affinity Purification ◽  
Protein A ◽  
High Sensitivity ◽  
Tandem Affinity Purification

Most cellular processes are carried out by a multitude of proteins that assemble into multimeric complexes. Thus a precise understanding of the biological pathways that control cellular events relies on the identification and on the biochemical characterization of the proteins involved in such multimeric assemblies. Advances in MS have made possible the identification of multisubunit protein complexes isolated from cell lysates with high sensitivity and accuracy, whereas the TAP (tandem affinity purification) methodology efficiently isolates native protein complexes from cells for proteomics analysis. TAP is a generic method based on the sequential utilization of two affinity tags to purify protein assemblies. During the first purification step, the Protein A moiety of the TAP tag is bound to IgG beads, and protein components associated with the TAP-tagged protein are retrieved by TEV (tobacco etch virus) protease cleavage. This enzyme is a sequence-specific protease cleaving a seven-amino-acid recognition site located between the first and second tags. In the second affinity step, the protein complex is immobilized to calmodulin-coated beads via the CBP (calmodulin-binding peptide) of the TAP tag. The CBP–calmodulin interaction is calcium-dependent and calcium-chelating agents are used in the second elution step to release the final protein complex preparation used for protein identification by MS. The TAP–MS approach has proven to efficiently permit the characterization of protein complexes from bacteria, yeast and mammalian cells, as well as from multicellular organisms such as Caenorhabditis elegans, Drosophila and mice.

scholarly journals Tandem affinity purification and identification of the human TSC1 protein complex

2010 ◽  
Vol 42 (4) ◽  
pp. 266-273 ◽  
Author(s):  
Longhua Guo ◽  
Wantao Ying ◽  
Jiyang Zhang ◽  
Yanzhi Yuan ◽  
Xiaohong Qian ◽  
...  
Keyword(s):  
Protein Complex ◽  
Affinity Purification ◽  
Tandem Affinity Purification

scholarly journals Tandem affinity purification in Drosophila: the advantages of the GS-TAP system

Fly ◽  
2008 ◽  
Vol 2 (4) ◽  
pp. 229-235 ◽  
Author(s):  
Phillip P. Kyriakakis ◽  
Marla Tipping ◽  
Louka Abed ◽  
Alexey Veraksa
Keyword(s):  
Affinity Purification ◽  
Tandem Affinity Purification

Tandem affinity purification of AtTERT reveals putative interaction partners of plant telomerase in vivo

PROTOPLASMA ◽  
2016 ◽  
Vol 254 (4) ◽  
pp. 1547-1562 ◽  
Author(s):  
Jana Majerská ◽  
Petra Procházková Schrumpfová ◽  
Ladislav Dokládal ◽  
Šárka Schořová ◽  
Karel Stejskal ◽  
...  
Keyword(s):  
Affinity Purification ◽  
Tandem Affinity Purification ◽  
Interaction Partners
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