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Membrane protein isolation


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#1 cancergeek

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Posted 16 October 2009 - 08:43 AM

Hi Folks,

I'm using a thermo fisher protein fractionation kit to isolate membrane proteins from my treated cells. In the protocol they mention the use of trypsin-edta to detach and collect cells prior to cell lysis. Wouldnt trypsin cleave off a number of membrane proteins? Or am I just being paranoid?

Thanks

#2 Holsten

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Posted 16 October 2009 - 03:39 PM

The amount of trypsin in Tr/EDTA solution is really small (0,05%), shouldn't do any harm in your case.
Alternatively, just to be on a safe side, you can use only EDTA (0.02%) in PBS, keep your flask on ice for 3-4 min and then transfer it to 37C. Hope this helps:-)




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