title: Purification of TAP-tagged proteins by two-step pull down from DT40 cells.
abstract: For proteomic analysis, protein purification from cell extracts is an important step. Since production of high quality antibody is time consuming and not guaranteed to be successful, expression of epitope-tag conjugated protein of interest followed by immunoprecipitation using anti-epitope-tag antibody is a common method for protein purification. Here we describe use of an epitope-tag called TAP (tandem affinity purification) in DT40, which consists of Protein A IgG-binding motif and calmodulin binding motif separated by TEV cleavage site. Tandem purification using two different epitopes should eliminate non-specific binding and help identifying physiological protein-protein associations.
authors.affiliation: Kawasaki Medical School, Department of Immunology and Molecular Genetics, 577 Matsushima, Kurashiki, Okayama 701-0192, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Kitao
authors.forename: Minoru
authors.initials: M
authors.lastname: Takata
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism*; D002460:Cell Line; D002645:Chickens; D002846:Chromatography, Affinity / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D011506:Proteins / Q000302:isolation & purification*
chemical_list: D011506:Proteins