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Ubiquilin / Dsk2 Antibody (131600) in WB
Western blot analysis of ubiquitin was performed by loading 30 µg of cell lysate of mouse thymocytes treated with 0 µM MG132 for 5 hr at 37C (lane 1) and cell lysate of mouse thymocytes treated with 20 µM MG132 for 5 hr at 37C (lane 2) in 1xSDS sample buffer and SeeBlue® Plus2 Pre-stained Protein Standard (Product # LC5925) onto a NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gels (Product # NP0324BOX). Proteins were transferred to nitrocellulose membrane. Membrane was blocked in 1xDPBS (Product # 14190136) containing 0.1% Tween 20 and 5% non-fat milk for one hour at room temperature. Ubiquitin was detected at numerous molecular weights using a Ubiquitin antibody (Product #13-1600) at a dilution of 1:1,000 in 1xDPBS (Product #14190136) containing 0.1% Tween 20 and 5% non-fat milk overnight at 4°C on a rocking platform, followed by a Goat anti-Mouse IgG-HRP secondary antibody at a dilution of 1:5,000 for one hour at room temperature. Chemiluminescent detection was performed using Immoblion Western and the ImageQuant LAS4000 imager. Data Courtesy of Dr. Zhi Wen at the Univeristy of Wisconsin, Madison.
