Hello. I am working with a membrane bound protein, and I've discovered, that after I dissolve and homogenize the tissue in RIPA buffer following by spin down, my protein of interest ends up in the pellet that is getting discarded.
How can I prepare the tissue and keep membrane bound proteins for western analysis?
Thank you
Engelbert Buxbaum
RIPA buffer contains detergent (SDS and cholate) and will solubilise membrane-bound proteins. Without knowledge of what protein you are interested in it is impossible to give specific advice, but you could try non-ionic caotropes like urea.
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