Figure 1. Western blot analysis of Glutaminase/GLS using anti-Glutaminase/GLS antibody (A01272-2). 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 50ug of sample under reducing conditions. Lane 1: human placenta tissue lysates, Lane 2: human U87 whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: monkey kidney tissue lysates, Lane 5: rat brain tissue lysates, Lane 6: rat kidney tissue lysates, Lane 7: rat heart tissue lysates. Lane 8: rat skeletal muscle tissue lysates. Lane 9: mouse brain tissue lysates. Lane 10: mouse kidney tissue lysates. Lane 11: mouse heart tissue lysates. Lane 12: mouse skeletal muscle tissue lysates. Lane 13: mouse Neuro-2a whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Glutaminase/GLS antigen affinity purified polyclonal antibody (Catalog # A01272-2) at 0.25 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-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for Glutaminase/GLS at approximately 65-73KD. The expected band size for Glutaminase/GLS is at 73KD.