RE:

From: Hawes, John W. (jhawes@iupui.edu)
Date: Fri Nov 17 2000 - 13:35:11 EST


Hi Mary,

Maybe you could pull out the his-tagged peptide using the MC-zip-tips
charged with nickel. These tips are used for metal chelation purification
of phosphopeptides, but I don't see any reason why they couldn't be used for
a his-tagged peptide. Has anyone else out there done that before?

The peptide may not be pure enough to sequence directly, but may be enough
to get a much more simplified chromotograph and sequence from there. ???

John Hawes
Biochemistry Biotechnology Facility
Indiana University School of Medicine
www.bbf.iu.edu

-----Original Message-----
From: Mary Bower [mailto:bowerma@purdue.edu]
Sent: Friday, November 17, 2000 10:52 AM
To: Recipients of ABRF List
Subject:

Dear colleagues,

I have a protein digest which has been separated by HPLC. My customer
suspects that there is a His tag on the N-terminal end of the protein, and
would like to sequence the N-terminal fragment. Does anyone know of a
paper that refers to a technique that enables you to identify such a
peptide from a mix?

Thanks in advance.

Mary Bower

Mary Bower
Laboratory for Macromolecular Structure
Dept. Biochemistry
Purdue Unversity
West Lafayette, IN 47907

voice: (765) 494-6540
fax (765) 494-7897



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