Title :
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
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;
Conference_Titel :
Remote Sensing, Environment and Transportation Engineering (RSETE), 2011 International Conference on
Conference_Location :
Nanjing, China
Print_ISBN :
978-1-4244-9172-8
DOI :
10.1109/RSETE.2011.5965772
