VWR Life Science RIPA Lysis Buffer, Biotechnology Grade

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	After cell harvesting, the pellets were washed three times with cold PBS, lysed with RIPA buffer containing protease inhibitors, and sonicated using a sonicator

Protocol tips
After cell harvesting, the pellets were washed three times with cold PBS, lysed with RIPA buffer containing protease inhibitors, and sonicated using a sonicator

Publication protocol

After cell harvesting, the pellets were washed three times with cold PBS, lysed with RIPA buffer (Ameresco, USA) containing protease inhibitors (Ameresco, USA), and sonicated using a sonicator. Protein concentrations were measured using a Pierce BCA protein assay kit (Thermo Scientific, USA). The cell lysates were subjected to polyacrylamide gel electrophoresis (PAGE, resolving gel 12% and stacking gel 4%). Then, the partitioned molecules were transferred to PVDF membrane by electrical blotting. The membranes were incubated with Odyssey blocking buffer for 1 h at room temperature, and then incubated with appropriate primary antibodies at 4oC overnight. After that, the membranes were incubated with accordant secondary antibodies at room temperature for 1 h. Band visualization and quantitation were carried out using a LI-COR Odyssey Imager (Li-COR, USA).

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