Anti- Gelsolin antibody, MA1044, IHC(P). IHC(P): Human Intestinal Cancer Tissue

Figure 3. Western blot analysis of Gelsolin using anti-Gelsolin antibody (MA1044). 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 Hela whole cell lysate, . Lane 2: human A549 whole cell lysate, . Lane 3: monkey COS-7 whole cell lysate, . Lane 4: human Caco-2 whole cell lysate, . Lane 5: human HepG2 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-Gelsolin antigen affinity purified monoclonal antibody (Catalog # MA1044) at 0.5 ug/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. A specific band was detected for Gelsolin at approximately 90KD. The expected band size for Gelsolin is at 86KD.