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. Author manuscript; available in PMC: 2015 Jul 9.
Published in final edited form as: Biol Chem. 2015 Mar;396(3):277–281. doi: 10.1515/hsz-2014-0191

Figure 4.

Figure 4

Immunoblot analysis demonstrating that the selected mAbs detect cathepsin B in lysates (0.68 μg DNA/sample) of MCF10-DCIS human breast cancer cells.

Lysates were run under reducing conditions on 12% SDS-PAGE gels at 50 mA and the separated proteins were transferred to nitrocellulose membranes. The membranes were incubated with the mAbs (1:500) for 2 h at room temperature and a secondary anti-mouse antibody (1:10 000). Detected cathepsin B was visualized through Western Lightening Plus ECL – Western Chemiluminescence Substrate. The 25/26 kDa band represents the heavy chain of the active two-chain form of cathepsin B and the 31 kDa band the active single-chain form of cathepsin B. Second step controls were negative and are not shown.