Purification and identification of HMGB1 secreted by liver cells and immune cells.
10.3969/j.issn.1672-7347.2011.11.006
- Author:
Meifang XIAO
1
;
Xiahong DAI
;
Rongrong ZHOU
;
Jianping LIU
;
Baoxin ZHANG
;
Shushan ZHAO
;
Xuegong FAN
Author Information
1. Department of Infectious Diseases, Xiangya Hospital, Central South University, Changsha 410008, China.
- Publication Type:Journal Article
- MeSH:
Cell Culture Techniques;
Electrophoresis, Polyacrylamide Gel;
methods;
HMGB1 Protein;
isolation & purification;
metabolism;
Hep G2 Cells;
Hepatocytes;
metabolism;
Humans;
Monocytes;
metabolism;
U937 Cells
- From:
Journal of Central South University(Medical Sciences)
2011;36(11):1065-1070
- CountryChina
- Language:Chinese
-
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.
