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Hakai Antibody (362800) in WB
Western blot analysis was performed on nuclear enriched extracts of HeLa (Lane 1), A549 (Lane 2), and U-2 OS (Lane 3). The blot was probed with Anti-Hakai Rabbit Polyclonal Antibody (Product # 36-2800, 2 ug/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 ug/ml, 1:2500 dilution). Approximately 70 and 75 kDa bands corresponding to the ubiquitinated form of Hakai were observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
