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?
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.
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.