Skip to main content
. Author manuscript; available in PMC: 2008 Mar 22.
Published in final edited form as: Biol Chem. 2006 Aug;387(8):1143–1146. doi: 10.1515/BC.2006.141

Figure 3. DPPI is processed normally in cathepsin L/S-/- mice.

Figure 3

Samples of 10 μg of total protein from cell lysates obtained from cathepsin L/S+/+, cathepsin L-/-, cathepsin S-/- or cathepsin L/S-/- BMMCs and purified dog DPPI were subjected to SDS-PAGE under reducing conditions and transferred to polyvinylidine difluoride membranes (Life Sciences Products, Boston, MA, USA). The membrane was washed with 50 mm Tris-HCl containing 0.5 m NaCl, 0.01% Tween-20 (TBS; pH 7.5), and incubated for 1 h in TBS containing either a 1:1000 dilution of rabbit anti-bovine DPPI (a gift from John Hoidal, University of Utah, Salt Lake City, USA) or monoclonal anti-β-actin antibody (Sigma-Aldrich). The membrane was then washed with TBS, incubated in TBS for 30 min containing a 1:2000 dilution of horseradish peroxidase-conjugated goat anti-rabbit IgG (New England Biolabs, Beverly, MA, USA) and washed again. Immunoreactivity was detected using the phototope-HRP detection kit (New England Biolabs). Note that bands of immunoreactive DPPI appear at similar sizes in both cathepsin L/S+/+ and cathepsin L/S-/- BMMC lysates.