Mitochondria Isolation

Ernesto Bustamante bustaman at AMAUTA.RCP.NET.PE
Fri Oct 18 11:10:03 EST 1996


At 12:24 PM 15/10/96 GMT, you wrote:
>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/
>
>

Dear friends:

I suggest you look up our method for isolating mitochondria from rat liver:

Bustamante,E. et al. (1977) Analytical Biochemistry 80:401-408 "A high-yield
preparative method for isolation of rat liver mitochondria".

This results in final mitochondrial yields of about 40 mg of mitochondrial
protein per gram wet weight of liver. These mitochondria are shown to be
functionally and structurally intact. 

A good general review can be found in Methods in Cell Biology 20: 411-481
(1978).

Good luck,

Ernesto
 


*****************************************************************************
Dr. Ernesto Bustamante                      
AB Chimica Laboratorios S.A.           Phone    51-1-224-3446
BelgaMedica S.A.                                Fax         51-1-224-2372
Av. Principal 592                                  bustaman at amauta.rcp.net.pe
San Isidro                                              71754.341 at compuserve.com
Lima 27,  Peru                		Ebustamant at aol.com
			AMICUS PLATO,  SED  MAGIS AMICA VERITAS
*****************************************************************************




More information about the Methods mailing list