NP-40 reduces contamination by endogenous biotinylated carboxylases during purification of biotin tagged nuclear proteins.
Protein Expr Purif
; 89(1): 80-3, 2013 May.
Article
em En
| MEDLINE
| ID: mdl-23500724
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.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Polietilenoglicóis
/
Biotina
/
Proteínas Nucleares
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article