Mitochondria Isolation

[Mick Partis]

Delvac at eijkman.go.id (Delvac Oceandy) wrote:
>I have problem in isolating mitochondria from mammalian tissues (e.g. liver,
>kidney, muscle)
>My problem is in the homogenization of the tissue in isolation buffer 
>If somebody has experience in isolating mitochondria, please tell me the
>best method to homogenize the tissue. 
>Which one is better, using motor-driven homogenizer or hand operated
>homogenizer, to get a good yield and purity of mitochondria ??
>
>Delvac Oceandy
>Eijkman Institute 
>Jakarta Indonesia
><delv at eijkman.go.id>
>
>

I used to homogenise by hand, and this will probably give you a better 
yield of intact mitochondria than using a motor.  More important 
however is the 'fit' of the teflon plunger in a Potter homogeniser.  If 
the fit is too  tight, you will inevitably break mitochondria either by 
shear forces, or by cavitation under the plunger.  How tight is too 
tight?  That will probably be different for the different tissues you are 
homogenising.  If you want intact 'coupled' mitochondria, you will have 
to sacrifice yield.

Mick
-- 
mick.partis at hri.ac.uk         Horticulture Research International
http://www.geocities.com/CapeCanaveral/1957/