TOP1 Antibody (435900) in WB

Western blot analysis was performed on nuclear enriched extracts (30 ug lysate) of HCT 116 (Lane 1), Hep G2 (Lane 2), Caco-2 (Lane 3) and PC-3 (Lane 4).The blots were probed with TOP1 Mouse monoclonal Antibody (Product# 435900, 2 ug/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 ug/ml, 1:4000 dilution). A 100 kDa band corresponding to TOP1 was observed across the cell lines tested. Apart from desired band, a 70 kDa band was observed in Hep G2 cell line representing the C terminal fragment of the enzyme. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), 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 following blocking with 5% skimmed milk. Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Western blot analysis was performed on nuclear enriched extracts (30 ug lysate) of HCT 116 (Lane 1), Hep G2 (Lane 2), Caco-2 (Lane 3) and PC-3 (Lane 4).The blots were probed with TOP1 Mouse monoclonal Antibody (Product# 435900, 2 ug/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 ug/ml, 1:4000 dilution). A 100 kDa band corresponding to TOP1 was observed across the cell lines tested. Apart from desired band, a 70 kDa band was observed in Hep G2 cell line representing the C terminal fragment of the enzyme. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), 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 following blocking with 5% skimmed milk. Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

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