GAPDH Antibody (6C5): sc-32233

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	Western blotting was carried out as described earlier (19). Briefly, the cells were lysed in RIPA buffer containing protease inhibitor and phosphatase inhibitor cocktails (Thermo Fisher Scientific). The nuclear protein was isolated according to the protocol provided by the Nuclear Protein Extraction Kit (Thermo Fisher Scientific). Then, equal amounts of protein was resolved on 10% SDS-PAGE and transferred to a PVDF membrane (Millipore). Membranes were incubated with primary antibody overnight at 4°C. Membranes were washed with TBST and incubated with horseradish peroxidase–conjugated secondary antibody. Proteins were visualized using ECL detection reagents (Beyotime Co.).

Protocol tips

Western blotting was carried out as described earlier (19). Briefly, the cells were lysed in RIPA buffer containing protease inhibitor and phosphatase inhibitor cocktails (Thermo Fisher Scientific). The nuclear protein was isolated according to the protocol provided by the Nuclear Protein Extraction Kit (Thermo Fisher Scientific). Then, equal amounts of protein was resolved on 10% SDS-PAGE and transferred to a PVDF membrane (Millipore). Membranes were incubated with primary antibody overnight at 4°C. Membranes were washed with TBST and incubated with horseradish peroxidase–conjugated secondary antibody. Proteins were visualized using ECL detection reagents (Beyotime Co.).

Publication protocol

Western blotting was carried out as described earlier (19). Briefly, the cells were lysed in RIPA buffer containing protease inhibitor and phosphatase inhibitor cocktails (Thermo Fisher Scientific). The nuclear protein was isolated according to the protocol provided by the Nuclear Protein Extraction Kit (Thermo Fisher Scientific). Then, equal amounts of protein was resolved on 10% SDS-PAGE and transferred to a PVDF membrane (Millipore). Membranes were incubated with primary antibody overnight at 4°C. Membranes were washed with TBST and incubated with horseradish peroxidase–conjugated secondary antibody. Proteins were visualized using ECL detection reagents (Beyotime Co.).

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