You can inject an inline 0.5um filtered sample in 6M GuCl (with reduction
alkylation reagents if you like) adjusted to have pH below 3 on to a reverse
phase column in TFA/acetonitrile with no ill effects to th e column and
excellent chromatogams (my true experience over ten years, mainly with
Zorbax SB, but also with Jupiter, Vydac, Supelco, Synchrom , according to
rough memory, not responsible for omissions!).
JOhn Holt
Aventis Pasteur
> -----Original Message-----
> From: Amanda Hall [SMTP:Amanda.Hall@newcastle.edu.au]
> Sent: Monday, March 05, 2001 12:31 AM
> 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