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Cell Lysis and Western Blotting

Cell lysis THP1

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#1 Eimear

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Posted 01 September 2011 - 04:16 AM

Hi

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!!

#2 GNANA

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Posted 01 September 2011 - 06:26 AM

I dont think RIPA would interfere your gel run!! post your RIPA composition and also tell how it interferes in the running of lysate...
I would prefer being perfectionist rather than a passionist in Research.

I always had an alternate hypothesis....





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