Rapid purification of 20S proteasome by ultrafiltration and ion-exchange membrane

Author

Li, Dong ; Guoqing ; Sun, Qing ; Lu, Xiaoshen ; Sui, Xinyu

Author_Institution

Department of biological science and engineering, School of chemical and biological engineering, University of science and technology Beijing, China

fYear

2011

fDate

24-26 June 2011

Firstpage

6194

Lastpage

6196

Abstract

Object: Rapid purification of 20S proteasome from fresh swine liver by ultrafiltration and ion-exchange membrane. Methods: 20S proteasome was first precipitated with ammonium, then separated by ultrafiltration membrane with cut molecular weight of 300 KD and 1000 KD, and finally separated by ion-exchange membrane. The purified fraction was verified by specific proteasome inhibitor epoxomicin, SDS-PAGE and western blotting. Results: The 20S proteasome was purified from fresh swine liver within 3 hours. The specific activity of purified 20S proteasome has about 100 fold of increase compared to the liver extracts. SDS-PAGE analysis and western blotting analysis with 20S proteasome antibody indicate that 20S proteasome was separated from other major proteins in the liver. Conclusion: A simple and rapid method to purify 20S proteasome from swine liver has been developed.

Keywords

Biomembranes; Inhibitors; Liver; Proteins; Purification; Sun; 20S proteasome; ion-exchange membrane; purification; ultrafiltration membrane;

fLanguage

English

Publisher

ieee

Conference_Titel

Remote Sensing, Environment and Transportation Engineering (RSETE), 2011 International Conference on

Conference_Location

Nanjing, China

Print_ISBN

978-1-4244-9172-8

Type

conf

DOI

10.1109/RSETE.2011.5965772

Filename

5965772