Hi,I am looking for some secreted proteins in the conditioned medium of cerebellar granule neurons. Since the amount can be small, I tried to use the Amicon Ultra-4 Centrifugal Filter Devices to concentrate the media before running them on SDS gel. But after doing that the gel would run funny - looks like partially dissolved. And I wasn't able to detect any clear bands. On the attached picture, the left 5 lanes are the CGN cell lysate, while the right side should be CM after concentration.Does anyone also use such devices for the same purpose? Or would you suggest I try other means of concentrating the CM?Thanks a whole bunch!
Did you tried speed vacume concentration?I guess that the problem that you had is that some other proteins or other lipids where also concentrated and thus the gel run funny.Maybe the way that I sugested whould help. You can always try to percipitate just proteins with TCA and clean the pellete with acetone.
Thank you very much! I'll give the speed vac a try.
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