MLKL Rabbit Monoclonal Antibody

Western blot analysis of MLKL using anti-MLKL antibody (orb866681). 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 PC-3 whole cell lysates, Lane 3: human A549 whole cell lysates, Lane 4: human MCF-7 whole cell lysates, Lane 5: human HepG2 whole cell lysates, Lane 6: human U87 whole cell lysates, Lane 7: human 293T 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-MLKL antigen affinity purified monoclonal antibody (Catalog # orb866681) at 1: 1000 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 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for MLKL at approximately 54 kDa. The expected band size for MLKL is at 54 kDa.