NP-40 reduces contamination by endogenous biotinylated carboxylases during purification of biotin tagged nuclear proteins
| Autor: | Jeroen Demmers, John Strouboulis, Dimitris N. Papageorgiou |
|---|---|
| Přispěvatelé: | Biochemistry |
| Rok vydání: | 2013 |
| Předmět: |
Cell Extracts
Cell Nucleus Lysis Chromatography Octoxynol Escherichia coli Proteins Biotin Nuclear Proteins Biology Contamination Mass spectrometry Polyethylene Glycols Repressor Proteins chemistry.chemical_compound Biochemistry Affinity chromatography chemistry Cytoplasm Biotinylation Carbon-Nitrogen Ligases Nuclear protein Biotechnology |
| Zdroj: | Protein Expression and Purification, 89(1), 80-83. Academic Press |
| ISSN: | 1046-5928 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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