HPLC -Reply
Henry Wolfe (hnwo@americas.nycomed.com)
Tue, 18 May 1999 08:37:10 -0600
I've never done it for phosphopeptides, but have found both CM and DEAE-based HPLC systems very useful. I have kept 25 % acetonitrile in both the eluting and loading solvent to sharpen the peaks and reduce sticking. Typically, I load in 10 mM sodium acetate, pH 5.5 and elute with 1o mM sodium acetate containing 1 M NaCl. I use a linear gradient, collect the maijor peak and run it right into a prep-LC after removing ~50 % of the ACN by rotovap.
good luck
Hank Wolfe
Nycomed Amersham
Wayne, Pa