Amanda,
In addition to worrying about whether the HOFo will keep your peptides from
binding properly, you have to remember that as soon as the acid runs away,
your peptide might crash (irreversibly) on your column. If it does, you
might be able to wash it off (saving your column but probably not your
peptide) with DMSO at a low flow rate. For such very hydrophobic peptides
you may want to try a C4 column, or even think about normal phase or CCD
purification.
Good luck!
Mark
-----Original Message-----
From: Amanda Hall [mailto:Amanda.Hall@newcastle.edu.au]
Sent: Sunday, March 18, 2001 10:37 PM
To: Recipients of ABRF List
Subject: Protsep:insoluble peptides
Hi everyone,
Continuing on from my question of a couple of weeks ago. I have a milk by
product "filter cake" that is highly insoluble. I need to get it into
solution so that I can separate the peptides contained within it by HPLC
and then sequence them.
I have managed to get the filter cake into solution by dissolving in 70%
formic acid, which I then diluted to 35%. My question is this- will 35%
formic acid affect (or even destroy) a C18 column or have any affect when
mixed with acetonitrile?
Thanks alot
Amanda
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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 - 10:20:50 EDT