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Capricy, and everybody: My guess is that the ionic strength around your protein was high enough to prevent the initial implant onto the column. Dilution lowered the ionic strength. Also, your buffer concentration is arguably a little too high. There are nice chromatography brochures available from www.vydac.com . Click on "Publications" then "RP Handbook and other brochures". The document about ion exchange is useful. Happy elution, Dan Anderson On Sun, 23 Oct 2005, capricy gao wrote: > Hi > > My protein of around 15mg/ml (0.3mM) was loaded to anion exchange column with > 25mM Tris.HCl, pH8.0, but it came out before the gradient started, never > sticking to the column. After I did the dilution by 10 to 15 fold, it showed > to bind to the column.Anybody has the similar experience and share some > explanations? > > Thanks, > > Capricy > > > --------------------------------- > Yahoo! FareChase - Search multiple travel sites in one click.
