Samples were separated by Novex NuPAGE SDS–PAGE system (Thermo Fisher Scientific) using 4–12% Bis-Tris in MES running buffer according to the manufacturer’s guidelines, and transferred to a 0.45 μm nitrocellulose membrane (Bio-Rad) by tank-blot wet transfer (Bio-Rad) at room temperature for 1 h. Blocking, primary antibody incubations (4 °C, 16 h) and secondary antibody incubations (room temperature, 1.5 h) were performed in 5% (w/v) milk (Maresi, Fixmilch instant milk powder) in PBS, containing 0.05% Tween-20 (Sigma-Aldrich). SMC4 was probed using a rabbit polyclonal antibody (Abcam, ab229213, GR3228108-5, 1:1,000). GAPDH was probed with a rabbit polyclonal antibody (Abcam, ab9485, GR3212164-2, 1:2,500). hKid was probed with a monoclonal rabbit antibody (Abcam, ab75783, GR129278-4, 1:1,000). KIF4A was probed with a recombinant rabbit antibody (Abcam, ab124903, GR96215-7, 1:1000). Horseradish peroxidase (HRP)-conjugated anti-mouse or anti-rabbit secondary antibodies (Bio-Rad, 1:10,000) were visualized using ECL Plus Western Blotting Substrate (Thermo Fisher Scientific) on a Bio-Rad ChemiDoc Imager operated by analysed using Bio-Red Image Lab v.6.0.1, which was also used for analysis.
Novex nupage sds page system
Manufactured by Thermo Fisher Scientific
The Novex NuPAGE SDS–PAGE system is a pre-cast polyacrylamide gel electrophoresis (SDS-PAGE) system designed for the separation and analysis of proteins. The system includes pre-cast gel cassettes, running buffers, and accessories for efficient and consistent protein separation.
Automatically generated - may contain errors
Lab products found in correlation
Ab124903, by Abcam (2 mentions)
Ab75783, by Abcam (2 mentions)
Ab229213, by Abcam (2 mentions)
Chemidoc imager, by Bio-Rad (2 mentions)
Tank blot wet transfer, by Bio-Rad (2 mentions)
Ecl plus western blotting substrate, by Thermo Fisher Scientific (2 mentions)
Tween 20, by Merck Group (2 mentions)
Ab9485, by Abcam (2 mentions)
0.45 μm nitrocellulose membrane, by Bio-Rad (1 mentions)
Anti mouse or anti rabbit secondary antibodies, by Bio-Rad (1 mentions)
0.45 m nitrocellulose membrane, by Bio-Rad (1 mentions)
2 protocols using novex nupage sds page system
1
Detailed SDS-PAGE Western Blotting Protocol
2
Western Blotting for Protein Analysis
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Samples were separated by Novex NuPAGE SDS–PAGE system (Thermo Fisher Scientific) using 4–12% Bis-Tris in MES running buffer according to the manufacturer’s guidelines, and transferred to a 0.45 µm nitrocellulose membrane (Bio-Rad) by tank-blot wet transfer (Bio-Rad) at room temperature for 1 h. Blocking, primary antibody incubations (4 °C, 16 h) and secondary antibody incubations (room temperature, 1.5 h) were performed in 5% (w/v) milk (Maresi, Fixmilch instant milk powder) in PBS, containing 0.05% Tween-20 (Sigma-Aldrich). SMC4 was probed using a rabbit polyclonal antibody (Abcam, ab229213, GR3228108-5, 1:1,000). GAPDH was probed with a rabbit polyclonal antibody (Abcam, ab9485, GR3212164-2, 1:2,500). hKid was probed with a monoclonal rabbit antibody (Abcam, ab75783, GR129278-4, 1:1,000). KIF4A was probed with a recombinant rabbit antibody (Abcam, ab124903, GR96215-7, 1:1000). Horseradish peroxidase (HRP)-conjugated anti-mouse or anti-rabbit secondary antibodies (Bio-Rad, 1:10,000) were visualized using ECL Plus Western Blotting Substrate (Thermo Fisher Scientific) on a Bio-Rad ChemiDoc Imager operated by analysed using Bio-Red Image Lab v.6.0.1, which was also used for analysis.
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