If you deglycosylate the protein, would it become susceptible to
proteinase K? Might be worth a shot.
Peter
On 29/08/2013 21:18, Ed Siefker wrote:
I am trying to extract nucleic acids from mucinous tumors and ascites.
These samples are predominately mucin2, which contains a proteinase
K resistant glycoprotein domain.
I've been using typical methods of extraction, namely proteinase K digestion
(which does make the lysate much less viscous), phenol extraction,
and ethanol precipitation. Unfortunately, no matter how many times
I extract with phenol, there is always material left at the interface, and
ethanol precipitation yeilds a pellet that is mostly protein.
There is a protease that digests mucin, EhCP5, but it is not commercially
available, and I'd like to avoid making my own recombinant proteins.
What alternative techniques should I be looking at?
-Ed
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