is it the same protein?

Ian McFarlane I.McFarlane at icrf.icnet.uk
Wed Mar 11 12:58:17 EST 1998


If you run SDS-PAGE with mercaptoethanol you should only get one band on
your Western. If this doesn't work, some protein complexes will resist the
treatment urea can be added to the sample buffer and gel. 

Ian Mc



In article <19980311131600.IAA08911 at ladder03.news.aol.com>, alpies at aol.com
(AlPiEs) wrote:

> We are studying a human protein that we suspect has a tendency to
aggregate in 
> vivo. After size exclusion chromatography and Western blot analysis of the
> fractions with polyclonal antibodies,there are two strong signals: one in the
> first fraction, and one in later fraction. Presumably the first fraction
> contains an aggregated form of the protein.  My question is: how do we show
> that the protein present in both fractions is the same? We do not want to rely
> on the antibodies only, and we cannot do microsequencing, the amounts are too
> low. Limited proteolysis is not agood idea either if it is the same protein in
> different conformations. Any suggestions, please?



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