title: Tandem affinity purification combined with mass spectrometry to identify components of protein complexes.
abstract: Most biological processes are governed by multiprotein complexes rather than individual proteins. Identification of protein complexes therefore is becoming increasingly important to gain a molecular understanding of cells and organisms. Mass spectrometry-based proteomics combined with affinity-tag-based protein purification is one of the most effective strategies to isolate and identify protein complexes. The development of tandem-affinity purification approaches has revolutionized proteomics experiments. These two-step affinity purification strategies allow rapid, effective purification of protein complexes and, at the same time, minimize background. Identification of even very low-abundant protein complexes with modern sensitive mass spectrometers has become routine. Here, we describe two general strategies for tandem-affinity purification followed by mass spectrometric identification of protein complexes.
authors: Department of Biological Chemistry, College of Medicine, University of California Irvine, Irvine, CA, USA.
authors: Peter
authors: P
authors: Kaiser
authors: David
authors: D
authors: Meierhofer
authors: Xiaorong
authors: X
authors: Wang
authors: Lan
authors: L
authors: Huang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D013058:Mass Spectrometry / Q000379:methods*; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D011506:Proteins