Application Notes: Western blot, 0.25-0.5 µg/ml, Human ELISA, 0.1-0.5 µg/ml, -. Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml
Western blot analysis of VCAM1 using anti-VCAM1 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 30 ug of sample under reducing conditions. Lane 1: human Hela whole cell lysates, Lane 2: human A549 whole cell lysates, Lane 3: human COLO320 whole cell lysates, Lane 4: human A431 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 rabbit anti-VCAM1 antigen affinity purified polyclonal antibody at 0.5 µg/mL overnight at 4C, 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 1:5000 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 VCAM1 at approximately 100 kDa. The expected band size for VCAM1 is at 81 kDa.
Western blot analysis of VCAM1 using anti-VCAM1 antibody.
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