MYOD1 Antibody / Myoblast determination protein 1 (FY13124)

Western blot analysis of MYOD1 using anti-MYOD1 antibody. Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. Lane 1: human RD whole cell lysates, Lane 2: rat heart tissue lysates, Lane 3: rat skeletal muscle tissue lysates, Lane 4: mouse heart tissue lysates, Lane 5: mouse skeletal muscle 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-MYOD1 antibody at 0.5 ug/ml overnight at 4oC, 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 was developed using an ECL Plus Western Blotting Substrate. MYOD1 antibody detects a dominant band at ~45 kDa in human RD cells and at ~40 kDa in mouse and rat tissues. Although the theoretical molecular weight is ~35 kDa, MYOD1 typically migrates slower due to phosphorylation and its basic DNA-binding domain. The weaker lower band in the human sample likely represents a dephosphorylated or partially cleaved form.