I want to resolve both membrane and cytoplasmic proteins from the same sample. In my hands, the membrane-bound protein is best resolved with NP40-containing buffer (150mM NaCl) while the cytoplasmic protein is best resolved with triton-x (300mM NaCl) containing buffer (as expected). so then I was thinking....why not mix both into one buffer.
Hi,
You can try, but your are going to get mixed micelles, and this can gives place to some unexpected results.
On the other hand,
a) salt concentration is also different . This could affect resolution
b) I do not understand why you need Triton-X (100?) to resolve cytoplasmic proteins. Cytoplasmic proteins are soluble, so detergent should not be necessary.
Good luck
Thank you! The cytoplasmic protein I'm trying to resolve is actually a small peptide that comes in both a soluble and insoluble form -- hence the detergent.
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