Anti-Talin is tested for Western Blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately ~270-272 kDa corresponding to the appropriate cell lysate or extract.
Western blot analysis of Talin using anti-Talin antibody. 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 30µg of sample under reducing conditions. Lane 1: human Daudi whole cell lysates, Lane 2: mouse NIH/3T3 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 affinity purified mouse anti-Talin antigen monoclonal antibody at 1 µg/mL overnight at 4C, 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 1:10,000 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 Talin at approximately 272 kDa. The expected band size for Talin is at 272 kDa.
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