Journal: Journal of Bacteriology
Article Title: Proteome and Transcriptome Analysis of the Virulence Genes Regulated by the VirR/VirS System in Clostridium perfringens
Figure Lengend Snippet: SDS-PAGE analysis of purified protease. Concentrated culture supernatant was applied to a benzamidine-Sepharose 6B column (2 ml) in 50 mM Tris-HCl (pH 8.0)-0.5 M NaCl. The retained enzyme was eluted by applying 0.1 M glycine-HCl (pH 3.0). Fractions of 1 ml were collected and monitored for enzyme activity. Enzyme activity was determined by using azocoll as a substrate. Fractions showing proteolytic activity were pooled. Protein concentration was determined by the Bradford method, and this fraction was subjected to SDS-PAGE analysis under reducing conditions.
Article Snippet: Concentrated supernatant solution was applied to a benzamidine-Sepharose 6B (Amersham Pharmacia Biotech United Kingdom, Ltd., Buckinghamshire, England) affinity column (bed volume, 2 ml) equilibrated with 50 mM Tris-HCl (pH 8.0) containing 0.5 M NaCl.
Techniques: SDS Page, Purification, Activity Assay, Protein Concentration