sonication of PBMC - (Feb/05/2001 )

sir, I am in desperate need of a protocol for the sonication of PBMC for animportant experiment, where I would ultimately do a Western Blot with the protein extract . Please forward any advise with this regard to my e-mail address.

PBMC isolation has to be carried out with the utmost care. the normal frequencies that can be used range from about 500-600 Hz. the pulses are sent at frequencies of around 1 minute each followed by three minutes of waiting period. the cells are suspended in saline to maintain osmolrtiy and as always ice is used for removing the heat generated.hope this helps you.please feel free to ask further questions.