Cell Lysis Buffer (10X)

Protocol tips

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	- Total proteins is extracted with Cell Lysis Buffer containing 1 mM phenylmethanesulfonyl fluoride (Sigma) on ice. The lysate is scraped and sonicated for 5 s by using an ultrasonic disruptor at output 1. The sonicate is centrifuged at 15,000 rpm for 10 min at 4°C.

Protocol tips
- Total proteins is extracted with Cell Lysis Buffer containing 1 mM phenylmethanesulfonyl fluoride (Sigma) on ice. The lysate is scraped and sonicated for 5 s by using an ultrasonic disruptor at output 1. The sonicate is centrifuged at 15,000 rpm for 10 min at 4°C.

Publication protocol

Total proteins from KC02-44D cells were extracted with Cell Lysis Buffer (Cell Signaling Technology, Beverly, MA) containing 1 mM phenylmethanesulfonyl fluoride (Sigma) on ice. The lysate was scraped and sonicated for 5 s by using an ultrasonic disruptor at output 1 (Tomy Digital Biology, Tokyo, Japan). The sonicate was centrifuged at 15,000 rpm for 10 min at 4°C, and the protein contents of the supernatant were measured using the Bradford protein assay (Bio-Rad, Hercules, CA). After the addition of lithium dodecyl sulfate sample buffer (Life Technologies, Grand Island, NY) containing 2-mercaptoethanol (Sigma), the samples were reduced at 70°C for 10 min.

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