about cell sonication - (Jul/21/2005 )

hi everyone, i want to destory rat hepatocyte of primary culture endoplasmic reticulum to obtain an enzyme by sonication(5*10e6cells/500ul homogenate buffer),can you tell me how long and how many times should i take?

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Hi there,
There is no set rule for sonication because there are many factors that influence the efficiently of the sonication, i.e. viscosity, amount, type of cells.

The basic protocol I use mammalian COS cells is to pulse 10 sec on and 30 sec off 4 times @ 40% amp with a microtip.

HTH

Jeff

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I sonicated mammalian HeLa cells and I did 3 times 10 sex with intervals of 30 sec.
Very important is to keep the tube with cell lysate always on ice.
Even during sonication tube should be on ice.

Good luck

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