[Purification and identification of HMGB1 secreted by liver cells and immune cells].
Zhong Nan Da Xue Xue Bao Yi Xue Ban
; 36(11): 1065-70, 2011 Nov.
Article
em Zh
| MEDLINE
| ID: mdl-22169720
ABSTRACT
OBJECTIVE:
To purify and identify HMGB1 secreted by liver cells HepG2 and immune cells U937.METHODS:
We cultured the liver cell lines HepG2 and immune cell lines U937, and stimulated them with HMGB1 (400 ng/mL) for 20 h. Then the supernatant was collected. Ultrafiltration centrifugation, CM-Sepharose cation, DEAE-Sepharose anion exchange chromatography, Sephadex G75-gel filtration chromatography, and immunoprecipitation were used for purification. The molecular weight and identity of HMGB1 was confirmed by SDS-PAGE and Western blot.RESULTS:
A sharp stained protein band with a molecular weight of about 26 kD was obtained by SDS-PAGE analysis and shown to be HMGB1 confirmed by Western blot.CONCLUSION:
High purified HMGB1 can be separated from these two cell lines.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Monócitos
/
Hepatócitos
/
Proteína HMGB1
Tipo de estudo:
Diagnostic_studies
Limite:
Humans
Idioma:
Zh
Ano de publicação:
2011
Tipo de documento:
Article