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Protein Expr Purif. 2013 May;89(1):80-3. doi: 10.1016/j.pep.2013.02.015. Epub 2013 Mar 7.

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

Author information

1
Division of Molecular Oncology, Biomedical Sciences Research Center Alexander Fleming, 34 Fleming Street, Vari GR-166 72, Greece.

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.

PMID:
23500724
DOI:
10.1016/j.pep.2013.02.015
[Indexed for MEDLINE]

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