2 replies to this topic

[anthonysinai]

Hello,
I am looking for a reliable protocol to osmotically swell mammalian liver mitochondria.  The purpose of this is to selectively protease treat the IMS while leaving intact mitoplasts.  Any assistance would be appreciated since most of the protocols I have found deal with yeast and fungal (neurospora) mitochondria.  
many thanks
Tony

[grzesiek]

Hello. Did you consider performing a selective lysis of the mitochondrial outer membrane with digitonin? Here's one reference : Proc Natl Acad Sci U S A 1998 Jan 6;95(1):144-9.
Regards, Grzegorz Kudla

[anthonysinai]

Thanks for the reference.  Unfortunately digitonin appears to be very inconsistent in our hands which is why we were hoping to get a good osmotic swelling protocol.  We found one in Methofs in Enzymol which we will be testing out soon.
Thanks anyway
Tony