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Cultured Cells Sample 1. Pipette proper volume of RIPA Lysis Buffer and mix thoroughly. 2-3 min advanced before use, add PMSF buffer to make its final concentration to1mM. 2. For adherent cells: wash sample with PBS, normal saline or serum-free culture medium to remove culture solution. Add proper volume of RIPA Lysis Buffer, then stroke with pipette until the buffer immerse cells completely. Shake slightly for 5-10 min. After lysis, centrifuge at 10,000-14,000 g for 10 min, then collect the supernatants and move on to the next step. >> Instruction for RIPA usage: for different sizes of cell culture plates <<