Cell lysis in detergent-free Hypotonic Lysis Buffer and the subsequent isolation of the crude cytoplasmic fraction can be completed in less than 30 minutes. The protocol is compatible with a variety of downstream applications including Western blotting and ELISA procedures as well as protein assays, which are sensitive to the presence of detergents in the lysis buffer. Simply, resuspend the cell pellets in Hypotonic Lysis Buffer and incubate on ice for 15 minutes. Disrupt the cells by homogenization and pellet the cellular debris. The supernatant contains the crude cytoplasmic extract.
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