Does ultrasonic probe sonication break up mitochondria or nuclear membrane? - (May/09/2015 )

Hello All,

Does ultrasonic probe sonication break up mitochondria or nuclear membrane of cultured cell?

I am confused about this for long time. The ultrasonic probe sonication is used in some paper to homonoginize cell lysate then centrifuge to isolate intact mitochondria (no breaking mitochondrial membrane). However many guides in google search also mention that the sonication can break every organelle membrane. Why is so different? Is it about the power setting or time period to sonicate cell on ice?

In addition, Does the sonication disassociate protein-protein interaction in CO-IP?

Thank you very much

It all depends on the sonication conditions (amplitude and time).  Different targets of extraction will require different settings.  And the generation of heat from the sonication can cause dissociation.  I am not sure about physical disruption, but that is one of many reasons for cross-linking in a ChIP.  Is there a reason for doing sonication, such as tissue extraction?  If not, I would suggest you use detergents to lyse your cells.

JDSBlueDevl on Tue May 12 04:07:18 2015 said:

It all depends on the sonication conditions (amplitude and time).  Different targets of extraction will require different settings.  And the generation of heat from the sonication can cause dissociation.  I am not sure about physical disruption, but that is one of many reasons for cross-linking in a ChIP.  Is there a reason for doing sonication, such as tissue extraction?  If not, I would suggest you use detergents to lyse your cells.

Thank you JDS. My goal is to use sonicator to homoginize the cell before doing nuclei and mitochondria isolation for Western blot analysis. Normally they will use douce homoginizer, but my lab doesn't have. Do you know what setting for the sonicator is to just homoginize the cell? I would like to try detergent as well, but mild detergent such as tritonX100 or NP40 seems to soulubilze mitochondria membrane, which I want to keep intact.