Re: another CNBr-cleavage question

Deb McMillen (mcmillen@morel.uoregon.edu)
Thu, 19 Feb 1998 14:22:40 -0800 (PST)

Henriette,
We use 0.1M HCl 30% acetonitrile, introducing the CNBr solubilized in
acetonitrile.
How are you going to purify your resulting hydrophobic peptides? Often by
mass spec we know that we have generated fragments that we want, but
finding them by RP HPLC is another story--sometimes they are too
hydrophobic to come off of a C18 or C4 column.
Deb McMillen
Institute of Molecular Biology
University of Oregon
Eugene OR

On Thu, 19 Feb 1998, Henriette Remmer wrote:

> Dear ABRFers:
>
> I want to do a CNBr-cleavage of a hydrophobic 140 kD protein, which is not
> soluble in aqueous solution. So I plan to add some acetonitrile to the 70%
> aqueous formic acid during reaction.
> Is this advisable or are there other (better) ways of doing a Br CN
> cleavage with an water insoluble protein?
>
> Thanks in advance for taking your time!
> Henriette
>
>
> Dr. Henriette A. Remmer
> Director of Protein Services
> University of IIllinois
> 311 Noyes Laboratory Box 62-1
> 505 S Mathews Ave
> Urbana, IL 61801
> phone: (217)333-4695
> fax: (217)244-1142
> email: remmer@staff.uiuc.edu
>
>