HI,
after sonication, and centrifuge with top speed for 30 min
at 4 degree,
there is a layer of lipids floating on the supernatant..
How you treat them?
Certainly I cannot ignore them , but no idea how they
produced and how to further proceed.
They seems definitively among the useless for IP .
thx,
Bus
0
2 replies to this topic
#2
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Posted 22 July 2009 - 12:24 PM
BioBus, on Jul 22 2009, 06:20 AM, said:
HI,
after sonication, and centrifuge with top speed for 30 min
at 4 degree,
there is a layer of lipids floating on the supernatant..
How you treat them?
Certainly I cannot ignore them , but no idea how they
produced and how to further proceed.
They seems definitively among the useless for IP .
thx,
Bus
after sonication, and centrifuge with top speed for 30 min
at 4 degree,
there is a layer of lipids floating on the supernatant..
How you treat them?
Certainly I cannot ignore them , but no idea how they
produced and how to further proceed.
They seems definitively among the useless for IP .
thx,
Bus
I usually just ignore the small pellicle that I get after centrifugation. If you think it might be a problem then you could always increase the ratio of sonication buffer to cells. Of course this means you have to sonicate a larger volume so, if you do this, you might want to split your lysate into two aliquots before sonicating.
#3
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Hmmm, I think it's working
Posted 22 July 2009 - 04:41 PM
Just remove the solution from under the lipid layer to a new tube. Or, conversely take the lipid layer off the tube and discard, it shouldn't contain anything of importance for your IP.
