1 reply to this topic

[cancergeek]

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

[Holsten]

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:-)