I'd appreciate feedback from IMAC ZipTip users about how often they
are successful in isolating phosphopeptides from in-gel digest
material. My experience is that these tips work fine with beta-casein
but getting results from 'real' samples is not so easy, especially at
the subpmol level. I think there has been comment before about the
need to clean up the sample by reverse phase ZipTip before attempting
IMAC but that doesn't seem to solve the problem. I can confirm by
examining the IMAC flow through that known phosphopeptides have been
removed, but recovering them seems to be in the lap of the gods. I am
interested in subsequent LCQ analysis by nanospray, not maldi.
Any help/advice/experiences welcome and what are opinions on Fe vs Cu
, Ni or Ga ions?.
Len
-- ********************************************************************* Dr Len C. Packman Assistant Director of Research Department of Biochemistry University of Cambridge 80 Tennis Court Road Cambridge, CB2 1GA, UKTel: +44 (1223) 333639 FAX: +44 (1223) 766002 e-mail: l.c.packman@bioc.cam.ac.uk
Protein and Nucleic Acid Chemistry Facility http://www.bio.cam.ac.uk/proj/adr/PNAC/pnac.html
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