How to avoid frothing in sonication - (Jan/09/2009 )
as title
I still can't consistently avoid frothing during sonication. It's somehow frustrating coz you know it may denature your protein. What factors do you think are important? types, shape and material of the container? how deep the probe is submerged?
when I google this someone say frothing actually is a sign of your bacterial cell being broken open. Is that true?
-kingswill-
I generally sonicate in RIPA buffer to avoid SDS. Also the sonicator should be deep inside the lysate.
Also substantial rest should be given after the pulse and also the pulse duration should be less .
-newarray-
the ChIP forum has a thread discussing foaming problem during sonication.
-bioforum-
QUOTE (newarray @ Jan 9 2009, 08:52 PM)
I generally sonicate in RIPA buffer to avoid SDS. Also the sonicator should be deep inside the lysate.
Also substantial rest should be given after the pulse and also the pulse duration should be less .
Also substantial rest should be given after the pulse and also the pulse duration should be less .
My buffer doesn't have SDS, but Triton
and i usually do "10s sonicate, 30s rest on ice" cycle
-kingswill-
QUOTE (bioforum @ Jan 10 2009, 02:51 AM)
the ChIP forum has a thread discussing foaming problem during sonication.
thx
-kingswill-