Osteopontin (Cleaved) Antibody (702184) in WB

Western blot analysis was performed on whole cell extracts (30 ug lysate) of U-2-OS (Lane 1), Hep G2 (Lane 2), MKN45 (Lane 3) and Mouse stomach (Lane 4). The blots were probed with ABfinity? Anti-Osteopontin Recombinant Rabbit Monoclonal Antibody (Product ## 702184, 1-2 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 35 kDa band corresponding to Osteopontin 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% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product ## WP20005).

Western blot analysis was performed on whole cell extracts (30 ug lysate) of U-2-OS (Lane 1), Hep G2 (Lane 2), MKN45 (Lane 3) and Mouse stomach (Lane 4). The blots were probed with ABfinity? Anti-Osteopontin Recombinant Rabbit Monoclonal Antibody (Product ## 702184, 1-2 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 35 kDa band corresponding to Osteopontin 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% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product ## WP20005).

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