3 replies to this topic

[asaren]

Does ultrasonic probe sonication break up mitochondria or nuclear membrane?

 

I read from some forum post that it can break up mitochondria or nuclear membrane with high frquency. What is the frquency? On contary, many papers use the sonication method to isolate intact mitochondria (no breaking mitochondrial membrane).

 

In addition, If I want to collect protein for CO-IP, Can I do the sonication with cell lysate?

 

Thank you very much

Edited by asaren, 09 May 2015 - 09:39 AM.

[Curtis]

That is one method, but read Afshin Samali's papers for rapid fractionation. And also Yi-Te Hsu's papers for CHAPS lysis buffer. In case you want to use other buffers (because I saw your other question as well) be careful with RIPA. It is a strong buffer and might change the conformation of the protein of interest.

[asaren]

That is one method, but read Afshin Samali's papers for rapid fractionation. And also Yi-Te Hsu's papers for CHAPS lysis buffer. In case you want to use other buffers (because I saw your other question as well) be careful with RIPA. It is a strong buffer and might change the conformation of the protein of interest.

 

Thank you Curtis. Just to make sure if I understand correctly. So using the ultrasonic probe soincation can break up both mitochondrial and nuclear membranes. Yes?

 

Moreover I've tried to search Afshin Samali's papers, but couldn't find his rapid fractionation method. Do you have the paper pubmed number? Thank you

[Curtis]

I have posted it here on this forum before. just search Bioforum and you might find it.