[list_product_images]IHC(P): Human Intestinal Cancer Tissue|IHC(P): Human Lung Cancer Tissue|IHC(P): Human Mammary Cancer Tissue|Figure 4. Western blot analysis of Cathepsin D using anti-Cathepsin D antibody (BM1577).
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 A549 whole cell lysate,

Lane 2: human placenta tissue lysate,

Lane 3: human PC-3 whole cell lysate,

Lane 4: human Hela whole cell lysate,

Lane 5: human Caco-2 whole cell 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 mouse anti-Cathepsin D antigen affinity purified monoclonal antibody (Catalog # BM1577) 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-mouse 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 # EK1001) with Tanon 5200 system. Specific bands were detected for Cathepsin D at approximately 33 and 48KD. The expected band size for Cathepsin D is at 45KD.[/list_product_images]