FW: [RASMB] Working with glycerol solutions

[Cole, James]

Dear Jo
In addition to the stuff that Tom mentioned you should also be aware that preferential hydration in glycerol-containing solutions increases the partial specific volume. This effect is pretty small: by averaging some published data from four proteins  [Gekko and Timasheff (1981) Biochemistry 20, 4667] we calculated a linear change in v-bar of: delta v-bar/% Glycerol = 3.33 +/- 0.38 x 10^-4 (ml/g)/%. Also, glycerol can enhance oligomerization of proteins. For example, the Kd for dimerization of cmv protease drops by about 10-fold in the presence of 20% glcyerol [Cole (1996) Biochemistry 35, 15601].

Regards,
Jim

-----Original Message-----
From: rasmb-bounces at server1.bbri.org [mailto:rasmb-bounces at server1.bbri.org] On Behalf Of pjgb at mrc-lmb.cam.ac.uk
Sent: Wednesday, February 15, 2006 7:58 AM
To: RASMB-Noticeboard
Subject: [RASMB] Working with glycerol solutions

Dear RASMBers,

Has anyone had experience of working with significant percentages of glycerol (10-25%), for either velocity or equilibrium runs?
Obviously the main problem is going to be the greatly increased viscosity, particularly at 5°C with 20°C less of a problem, but what other snags are there?  Also, does the theoretical reduced diffusion, at these high viscosities, work out in practice so that the boundaries are no broader after spinning for the extra time needed?

Jo

P.J.G. Butler,
MRC Laboratory of Molecular Biology,
Hills Road, Cambridge, CB2 2QH, UK.
Tel. +44 (0)1223 402296
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