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Cell Lysis and Western Blotting - (Sep/01/2011 )


I need to lyse THP1 monocytes and then use western blotting to quantify the levels of certain proteins in the lysate. I tried lysing the cells with RIPA buffer, but the high detergent concentration was incompatable with running a gel. I then tried Lamberts Break buffer, but the total protein concentration of my lysare was extremely low (i.e. the cells didnt lyse).

Does anyone have any suggestions ass to how I can successfully lyse THP1s in a buffer that is compatable with SDS PAGE??

Thank you in advance!!


I dont think RIPA would interfere your gel run!! post your RIPA composition and also tell how it interferes in the running of lysate...