Purification of TAP-tagged proteins by two-step pull down from DT40 cells.

Hiroyuki Kitao, Minoru Takata

Research output: Contribution to journalArticle

3 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)409-413
Number of pages5
JournalSub-cellular biochemistry
Volume40
Publication statusPublished - Jan 1 2006

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Purification
Epitopes
Proteins
Antibodies
Staphylococcal Protein A
Calmodulin
Cell Extracts
Immunoprecipitation
Proteomics
Immunoglobulin G

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology
  • Cancer Research

Cite this

Purification of TAP-tagged proteins by two-step pull down from DT40 cells. / Kitao, Hiroyuki; Takata, Minoru.

In: Sub-cellular biochemistry, Vol. 40, 01.01.2006, p. 409-413.

Research output: Contribution to journalArticle

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