Figure 1. Western blot analysis of Grp75 using anti-Grp75 antibody (M02561-2). Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human HepG2 whole cell lysates, Lane 3: rat brain tissue lysates, Lane 4: rat lung tissue lysates, Lane 5: rat PC-12 whole cell lysates, Lane 6: mouse brain tissue lysates, Lane 7: mouse lung tissue lysates, Lane 8: mouse RAW264.7 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Grp75 antigen affinity purified monoclonal antibody (Catalog # M02561-2) at 0.5 microg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for Grp75 at approximately 74 kDa. The expected band size for Grp75 is at 74 kDa.