Western blot analysis of Bax using anti-Bax antibody (PA1013-1). 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: Rat Testis Tissue Lysate Lane 2: Rat Kidney Tissue Lysate Lane 3: Rat Brain Tissue Lysate Lane 4: Rat Ovary Tissue Lysate Lane 5: HELA Cell Lysate Lane 6: MM231 Cell Lysate Lane 7: A549 Cell Lysate Lane 8: JURKAT Cell Lysate Lane 9: Human Placenta Tissue Lysate 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-Bax antigen affinity purified polyclonal antibody (Catalog # PA1013-1) at 0.5 μg/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 Bax at approximately 21&28KD. The expected band size for Bax is at 21KD.