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samples preparation repo preparation prepare reaper or np-40 buffer supplemented with fresh protease and phosphatase inhibitors prepare when needed calculate the total volume of Reaper buffer before processing the samples tissue processing cut the tissues into pieces on ice as quickly as possible to prevent degradation by proteases add 200 to 300 microliters of ice-cold Reaper lysis buffer for about five milligrams of tissue samples and homogenized on ice you centrifuge at 12,000 rpm for five minutes at four degrees centigrade then take the supernatant adherence cell processing take out the culture medium and wash it with ice-cold PBS you add a hundred and fifty microliters of prepared lysis buffer for every five million cells then crack the mixture on ice for five to ten minutes and mix it with a scraper take the disrupted sample into an EP tube centrifuge at 12,000 rpm for five minutes at four degrees centigrade then take the supernatant suspension cell processing transfer the cells
