Protein Isolation

Ali McBride amcbride at
Tue Feb 25 15:02:19 EST 1997

Fellow Netters,
		I have a question on how to isolate proteins using a sepharose A
column, that is conjugated to a unknown Ab.  This antibody is a
monoclonal Ab, as well as being classified as an IgG1 subtype.  I want
to know what you would recommend in terms of protocols, or papers that
might help me elude the answer to the problem(No Pun Intended).  I have
used the protocol from Harlow et al., directly.  I tried to elute with a
low pH glycine as well as MgCl2.  I tried to elute the protein, however,
all I found on my coomassie gel were bands correlating to heavy chain
and light chains of the Ab.  
If you have any suggestions I would greatly appreciate an email.

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