Separation of phospho- and dephospho- protein

Joseph Fernandez (fernaj@rockvax.rockefeller.edu)
Tue, 17 Nov 1998 16:28:03 -0500 (EST)

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What is the best method for seperation of a phospho-protein from its
dephospho-protein counterpart following in incomplete in vitro
phosphorylation reaction (stoichiometry of about 0.3). My septigenerian
boss suggested a Mercury column. I am trying an Fe3+-IDA (metal chelating
column). This column has been shown to selectivly bind phosho-amino acids
at lower pH. It isn't working for my protein. Are there other methods of
seperation of prepartive amounts of phospho-protein? Thank you imensly.

James Bibb
c/o Joe Fernandez

Joseph Fernandez
Associate Director
The Rockefeller university
Protein/DNA Technology Center
1230 York Ave. New York, NY 10021
Phone: (212)-327-8869
FAX : (212)-327-8620
email: fernaj@rockvax.rockefeller.edu
Lab Web Page: http:\\pdtc.rockefeller.edu

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