Amanda,
You might be able to get it all dissolved into one solution. Try
differing salt and pH, urea, GuHCl, etc. are the traditional methods. Since
your not interested in bio. activity, I would try conc. TFA or formic acid,
but be aware your HPLC column won't last long with samples injected with
these conc. solvents. Also try dissolving less protein/peptide (i.e.,
having a diluted protein mixture), because it doesn't take much to sequence
them.
David
David T. Chin, Ph.D.
Director
Protein Biotechnology Core Facility
Univ. of Missouri - Columbia
-----Original Message-----
From: Amanda Hall [mailto:Amanda.Hall@newcastle.edu.au]
Sent: Sunday, March 04, 2001 11:31 PM
To: Recipients of ABRF List
Subject: protseq: insoluble protein
Hi everyone,
I have received a sample that is basically a by-product of a casein
digestion, I think. It is currently in the form of a solid "filter cake" and
is supposed to be insoluble in water. The client wants me to be able to
sequence the peptides that are contained within it.
therefore I need suggestions as to ways to make this filter cake soluble so
that I can separate the peptides on a HPLC and then collect the peptides for
sequencing.Since I want to separate on a C18 column I can't dissolve in any
of the conventional solvents.
I have heaps of sample to try out your suggestions
thanks in advance
Amanda
|\ _,,,---,,_
zz /,`.-'`' -. ;-;;,_
|,4- ) )-,_..;\ ( `'-'
'---''(_/--' `-'\_)
Amanda Hall-Griffin
Professional Officer
Newcastle Protein
University of Newcastle
Ph (02) 4921-7299
Fax (02) 4921-6903
This archive was generated by hypermail 2b29 : Mon Apr 02 2001 - 10:20:49 EDT