Anti gpnmb antibody

Liver tissues from mice were lysed with T-PER tissue protein extraction reagent (Pierce, Rockford, IL, USA) supplemented with protease inhibitor (Roche Diagnostics, Basel, Switzerland). Equal amounts of cell lysates (10 μg) were separated on 10% SDS polyacrylamide gels (SDS-PAGE) and electroblotted onto polyvinylidene fluoride membranes. The blots were probed with antibodies specific for anti-Gpnmb antibody (R&D Systems), β-actin (SantaCruz Biotechnology, Inc., Dallas, Texas, USA). After incubating the membrane with horseradish peroxidase−conjugated secondary antibodies, reactivity was visualized using electrogenerated chemiluminescence Western blotting detection reagents (GE Healthcare, Tokyo, Japan) and a Chemi Doc XRS-J digital densitometer (Bio-Rad Laboratories, Hercules, CA, USA).