RIPA Buffer

Protocol tips

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	- Extraction of protein from cells was performed with radioimmunoprecipitation assay buffer lysis buffer according to manufacturer's protocol.

Protocol tips
- Extraction of protein from cells was performed with radioimmunoprecipitation assay buffer lysis buffer according to manufacturer's protocol.

Publication protocol

Extraction of protein from cells was performed with radioimmunoprecipitation assay buffer lysis buffer (Sigma), and concentrations of the extracted proteins were measured by a bicinchoninic acid protein assay kit (Beyotime, Beijing, China). The extracted proteins (30 µg for each lane) were separated using 10% SDS‐PAGE followed by transferring onto the polyvinylidene difluoride membranes (Millipore, Bedford, USA). The transferred membrane was then incubated with PBS in Tween 20 containing 5% non‐fat milk at room temperature for 1 hour. The membrane was then incubated with primary antibodies at 4°C overnight. After incubation with primary antibodies, the membrane was then incubated with relevant secondary antibodies conjugated with horseradish peroxidase (Santa Cruz Biotechnology) at room temperature for 2 hours. The western blot bands on the membrane were detected by the enhanced chemiluminescence reagents (Sigma).

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for RIPA Buffer below.

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