From: Jo Butler <pjgb@mrc-lmb.cam.ac.uk>
  To  : Andrew Leech , RASMB@bbri.org
  Date: Fri, 17 Dec 1999 16:15:00 +0000

Re: Anaerobic use and C-filled Epon

Andrew,

We have done some work with a reduced cytochrome b, where the oxidation
state of the Fe is crucial.  In this case we pretreated the assembled cells
by several cycles of vacuum and then argon.  After that we could quickly
fill them in the open lab, taking the reduced cytochrome through the septum
of a vial with an HPLC injection syringe and introducing it into the cell
sector directly, then closing the sector with the insert and screw asap.

Maybe we are even more paranoid than you, but I do know that this resulted
in spectra corresponding to the fully reduced cytrochrome after several
days of an equilibrium run.

Jo Butler

--On Fri, Dec 17, 1999 3:47 pm +0000 Andrew Leech <A.Leech@uea.ac.uk> wrote:

> 
> I would be interested in hints and tips about ultracentrifugation
> of oxygen sensitive (protein) samples. I am worried that residual
> O2 might stick to the surface of the charcoal-filled Epon cells.
> My current idea is to store the assembled cells in a glove box
> for some period then fill them, then transfer out to the machine
> (which is some distance away).
> 
> Am I being paranoid, or are there techniques to minimise oxygen
> contamination during cell filling and run setups? Any advice would
> be greatly appreciated.
> 
> Andrew
> 
> ===================================================================
> 
> Dr Andrew Leech                  *Tel +44 (0) 1603 592758
> Scientific Officer               *Fax +44 (0) 1603 592250
> School of Biological Sciences    *Email a.leech@uea.ac.uk
> University of East Anglia        *
> Norwich NR4 7TJ,  U.K.           *
> 



P.J.G. Butler,
MRC Laboratory of Molecular Biology,
Hills Road,
Cambridge, CB2 2QH,
UK.