ubiquitin and sds

Mr. G. Morley gmorley at hgmp.mrc.ac.uk
Thu Jul 4 10:27:49 EST 1996

Dear all,
          just a quick note to ask about precipitating
ubiquitin bound proteins.I noticed in the literature that
groups who are working on ubiquitin-protein complexes lyse
the cells with buffer that does not contain SDS and then precipitate 
the solution with W.G.A sepharose (not with an antibody against
the receptor they are studying). Is this because the contitions
required to seperate the protein-ubiquitin complex from
the Protein A/G beads (used in a normal antibody ppt) also releases
the ubiquitin from the protein?... When I performed a antiubiquitin
western after ppt my receptor with antibody I noticed dark
bands much lower down the gel than I expected...
If anyone could let me know this would be appreciated!
Thanks in advance..
                     Gary Morley (gmorley at rpms.ac.uk)

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