Does anyone know of a chemical lysis method for total protein isolation from coelomocytes? I'm currently using physical disruption in the presence of detergent. This gets me a fair amount of my protein of interest, but I am looking for possible alternatives.
If you don't care about protein activity you can try TRIzol.
Does isolating with Trizol require mechanical disruption beforehand (that's the only way I've seen)? My concern is that I might sheer some of the proteins during physical disruption (I currently use sonication, and know this happens sometimes), so I'm looking for methods that are primarily chemical disruption.
You would probably have to vortex the sample.
If that is too harsh, then maybe osmotic lysis?
Thanks! I'll have to give this a try next time I do a protein isolation. Vortexing should be fine - when I sonicate, I usually don't get too much protein degradation (well, at least of my protein of interest) - it's the occasional over-soication that worries me with my current method
Hi. Yes, sonicators used to bust open bacteria can be quite intense. Plus they get hot. A sonicating waterbath might be an alternative.
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