seems very hard to re-dissolve the sample prepared in this way, and is that the whole protein or some special kind of the protein hardly soluble ?
so sorry, i foget to say that the sample is for 2DE, and I want to know the difference between two samples, one is control the other is treated with stress or disease which make the sample even more complex and even more hardly to soluble with lysis buffer(7M urea 2M thiourea 4% CHAPS 5mm TBP)
i am wondering.it is the difference in solubility of the two samples lead to the difference in 2DE gel, or the protein components as i anticipated?
SDS should be able to dissolve pellet. if not sonication is best possible answer. i would suggest to add protease inhibitor cocktail (or PMSF) for sonication, and not to add SDS if ur heading for sonication.
SDS is incompatible with 2DE,
my sonicating probe is about 1CM in diameter, my protein sollution is no more than 1 ml and i am also worring about the heat caused by sonication, I mean the effect of ultrasonic cavitation. The protein should subject to MS identification, so who know what will happen after sonification.
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