NP-40 reduces contamination by endogenous biotinylated carboxylases during purification of biotin tagged nuclear proteins

Dimitris N Papageorgiou, Jeroen Demmers, John Strouboulis

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

We describe here a simple procedure for greatly reducing contamination of nuclear extracts by naturally biotinylated cytoplasmic carboxylases, which represent a major source of non-specific background when employing BirA-mediated biotinylation tagging for the purification and characterization of nuclear protein complexes by mass spectrometry. We show that the use of 0.5% of the non-ionic detergent Nonidet-40 (NP-40) during cell lysis and nuclei isolation is sufficient to practically eliminate contamination of nuclear extracts by carboxylases and to greatly reduce background signals in downstream mass spectrometric analyses.

Original languageEnglish
Pages (from-to)80-3
Number of pages4
JournalProtein Expression and Purification
Volume89
Issue number1
DOIs
Publication statusPublished - May 2013

Keywords

  • Biotin/chemistry
  • Biotinylation
  • Carbon-Nitrogen Ligases/chemistry
  • Cell Extracts/chemistry
  • Cell Nucleus/chemistry
  • Escherichia coli Proteins/chemistry
  • Nuclear Proteins/isolation & purification
  • Octoxynol
  • Polyethylene Glycols/chemistry
  • Repressor Proteins/chemistry

Fingerprint

Dive into the research topics of 'NP-40 reduces contamination by endogenous biotinylated carboxylases during purification of biotin tagged nuclear proteins'. Together they form a unique fingerprint.

Cite this