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SMAD4 Antibody (701682) in WB
Western blot analysis was performed on whole cell extracts (30 ug lysate) of A549 (Lane 1), A549 serum starved for 4 hours (Lane 2), PC -3 (Lane 3), PC-3 serum starved for 4 hours (Lane 4) .The blots were probed with ABfinity? Smad4 Recombinant Rabbit Monoclonal Antibody (Product# 701682 0.5-2 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). A clear 60 kDa band corresponding to Smad4 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), 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® Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody following blocking with 5%skimmedmilk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
