Figure 1. Western blot analysis of ACAT2 using anti-ACAT2 antibody (MBS1755206).

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 HepG2 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human Jurkat whole cell lysates,
Lane 4: human K562 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat RH35 whole cell lysates,
Lane 7: mouse liver tissue 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 rabbit anti-ACAT2 antigen affinity purified polyclonal antibody (#MBS1755206) at 0.25 ug/mL overnight at 4 degree 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 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for ACAT2 at approximately 41 kDa. The expected band size for ACAT2 is at 41 kDa.