[RASMB] more on dithionite and Hb

[Tom Laue]

Hi-
The epon centerpieces do leach O2, and soaking them, then storing them 
in an N2 or Ar atmosphere is necessary. I recall, too, that he was 
concerned about the Al-filled epon, though properly passivated Al-filled 
centerpieces should be no more reactive than charcoal-filled 
centerpieces. Though this probably does not apply to your work, KelF 
centerpieces are even worse when it comes to O2 leaching.
Best wishes,
Tom

John Philo wrote:
> Regarding handling oxygen-sensitive materials in the centrifuge, first I
> agree fully with Jack Kornblatt's statements about dithionite and had said
> something similar to Mitra privately yesterday, and recommended he try an
> enzymatic oxygen scavenging recipe I used for Hb kinetics studies in the
> past. 
>
> Second, one additional point is that Todd Schuster told me years ago that
> the Epon centerpieces are a significant source of oxygen. I believe the
> old-timers would soak the centerpieces in dithionite solutions for several
> days to deplete that oxygen and then the need for oxygen scavenging capacity
> is much reduced.
>
> John 
>
> -----Original Message-----
> From: rasmb-bounces at rasmb.bbri.org [mailto:rasmb-bounces at rasmb.bbri.org] On
> Behalf Of Jack Kornblatt
> Sent: Thursday, February 05, 2009 8:04 AM
> To: RASMB
> Subject: [RASMB] more on dithionite and Hb
>
> Hello Mitra
> Chance, I think, once described dithionite as man's worst enemy. the
> concentration that you are using is far in excess of what is need to keep
> your solutions anaerobic. The reaction products of dithionite are too
> numerous to list even if I could remember them. If you degas your solutions
> just before loading and then add dithionite to 1 mM this should give you the
> desired "low" oxygen.
> Is it really necessary to keep dithionite as low as possible? I have little
> experience with Hb but if we use dithionite and cytochrome c oxidase at 11
> mM there are sufficient biproducts generated that interpreting data is very
> difficult
>
> best
> jack kornblatt
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-- 
Department of Biochemistry and Molecular Biology
University of New Hampshire
Durham, NH 03824-3544
Phone: 603-862-2459
FAX:   603-862-0031
E-mail: Tom.Laue at unh.edu
www.bitc.unh.edu
www.camis.unh.edu