FIG. 5.

Analysis of protease II by SDS-PAGE, Western blot, and 2D electrophoresis. (A) SDS-PAGE and Western blot analysis of purified protease II. Lane 1 contains the Mono Q fraction showing purified protease II. Protein (4.4 μg) was applied to a SDS-10% PAGE gel. The electrophoretic run was performed at 100 V, and the gel was subsequently silver stained. Lane 2 shows the results of Western blot analysis of 2 μg of purified fraction using antibodies raised against Prot.II-29 peptide as the primary antibody. (B) 2D analysis of purified protease II. A concentrated sample of protease II was applied to a 7-cm ready strip gel to perform the first-dimension step. The 2D step was performed in an SDS-10% polyacrylamide gel that was silver stained. IEF, isoelectric focusing.