Michael,
Put the resin into a fritted vessel that you can
agitate, or stir in as well as filter through. Do not
use a magnetic stirrer as you'll grind up the resin
clogging your filter frits, etc.
First swell the resin in DMF about 10 minutes each,
three times.
Add 25% Piperidine in DMF about ten volumes, with
respect to the weight of the resin, and let mix for
ten minutes. Repeat once more.
Wash the resin 4 times ten volumes DMF 2 minutes each,
To acylate (I'm assuming the N-terminus?) add 20%
Acetic anhydride in DMF and let mix for about 20
minutes. Wash 2x DMF as above.
Rinse the resin 2 x Methanol ten minutes each, and dry
in vacuo.
As far as cleavage, look at the sequence, go get a
NovaBiochem catalog and look in the back for the best
cocktail for your sequence and follow those directions
with one caveat...prior to precipitation of the
peptide, reduce the cleavage cocktail volume in vacuo
at NGT 40C to about 10-20% of original volume, then
add to the ether.
Hope this helps.
Regards,
Steven Johnson, B.S. Chem.
Research Associate
Process Development Chemistry
Biomeasure, Inc. Milford, MA
--- Michael Moore <mmoore@uclink4.berkeley.edu> wrote:
> A student brought a resin bound peptide that they
> want cleaved and acetylated.
> They're not sure if the final fmoc group was
> removed. Can someone suggest
> manual procedure for removing final fmoc and
> acetylation prior to cleavage.
>
> Thanks in advance
>
> Michael
>
>
__________________________________________________
Do You Yahoo!?
Get Yahoo! Mail - Free email you can access from anywhere!
http://mail.yahoo.com/
This archive was generated by hypermail 2b29 : Mon Oct 23 2000 - 10:23:17 EDT