why glycerol in lysis buffer?
Dr Engelbert Buxbaum
engelbert_buxbaum at hotmail.com
Wed Nov 10 09:46:26 EST 2004
Ira Bell wrote:
> What is is the role of glycerol in the lysis buffer used for
> immunoprecipitation (mammalian proteins)?
The effect of osmolytes like glycerol on proteins has been discussed for
a long time, but is still not fully understood.
Clear is that the inclusion of osmolytes reduces the concentration of
available water in the solution, which the proteins need as "grease" for
conformational movements. Thus osmolytes stabilise proteins in a compact
conformation, resulting in higher denaturation temperatures, better
crystallisation and lower enzymatic activity.
Note that this effect is shown by compounds which have a high affinity
for water, other compounds may interact preferentially with the
proteins, leading to denaturation. This is the basis of the well known
Hofmeister series for ions.
High concentrations of osmolytes will prevent bacterial growth and
protect proteins during freezing by preventing ice crystal formation.
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