Cytokeratin 5 Mouse Monoclonal Antibody

Western blot analysis of Cytokeratin 5 using anti- Cytokeratin 5 antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sampleunder reducing conditions. Lane 1: human A431 whole cell lysates Lane 2: human SH-SY5Y whole cell lysates Lane 3: human Hacat 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- Cytokeratin 5 antigen affinity purified monoclonal antibody  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 with Tanon 5200 system. A specific band was detected for Cytokeratin 5 at approximately 62 kDa. The expected band size for Cytokeratin 5 is at 62 kDa.