Fetal calf sera can distort cell-based luminescent proteasome assays through heat-resistant chymotrypsin-like activity.
Anal Biochem
; 471: 23-5, 2015 Feb 15.
Article
em En
| MEDLINE
| ID: mdl-25447494
ABSTRACT
Luminescence-based proteasome activity assays use specific substrates that are supposed to be cleaved by cellular proteasome activity leading to luciferase substrates. Usually, control wells containing cell culture medium supplemented with antibiotics and fetal calf serum are used as background. Using the Proteasome-Glo chymotrypsin-like cell-based assay from Promega, we show here that fetal calf sera from different manufacturers contain heat-resistant, bortezomib-inhibitable, chymotrypsin-like activities that can interfere with proteasome activity assays. These data strongly recommend the use of pure phosphate-buffered saline (PBS) or serum-free medium during proteasome activity assays to diminish background luminescence and, thus, to obtain reliable results.
Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Quimotripsina
/
Artefatos
/
Soro
/
Complexo de Endopeptidases do Proteassoma
/
Ensaios Enzimáticos
/
Medições Luminescentes
Limite:
Animals
Idioma:
En
Revista:
Anal Biochem
Ano de publicação:
2015
Tipo de documento:
Article
País de afiliação:
Alemanha