Journal: Infection and Immunity
Article Title: Description of Staphylococcus Serine Protease ( ssp ) Operon in Staphylococcus aureus and Nonpolar Inactivation of sspA -Encoded Serine Protease
doi: 10.1128/iai.69.1.159-169.2001
Figure Lengend Snippet: FIG. 3. SDS-PAGE (A) and Western immunoblot (B) of secreted proteins from S. aureus RN6390 and isogenic derivatives. The molec- ular masses of protein standards are indicated to the left of each gel. (A) Lane 1, S. aureus RN6390; lane 2, SP6391; lane 3, SP6912; lane 4, RN6911. The amount of protein loaded represents the equivalent of 2.3 OD600 units (lanes 1 and 2) or 6.8 OD600 units (lanes 3 and 4) of original culture. Secreted proteins are visualized by staining with Coo- massie blue dye. (B) Purified V8 protease (60 ng) from S. aureus L530 (lane 1) or supernatant corresponding to 0.03 OD600 units of stationary phase culture from S. aureus RN6390 (lane 2), SP6391 (lane 3), SP6912 (lane 4), or RN6911 (lane 5) was subjected to SDS-PAGE (10% [wtzvol21] polyacrylamide), transferred to Immobilon-P membrane, and probed with blot-purified polyclonal anti-V8 protease antibody.
Article Snippet: For zymography, culture supernatants were subjected to SDSPAGE using 12% (wtzvol21) acrylamide Zymogram Ready Gels (Bio-Rad, Hercules, Calif.) containing casein or gelatin (1 mgzml21).
Techniques: SDS Page, Western Blot, Staining, Membrane