Tris buffer and the Biorad model 491 Preparative Electrophoresis

Dr. Duncan Clark Duncan at nospam.demon.co.uk
Mon Oct 18 08:01:55 EST 1999


In article <13C84303495 at rose.le.ac.uk>, Dr P.P.E. Freestone
<ppef1 at leicester.ac.uk> writes
>Hello, 
>We are using a Biorad model 491 Preparative Electrophoresis cell to
>purify collagen.  To optimise our purification, we'd like to collect
>discrete peaks by connecting the cell tube outlet to a flow cell. 
>However, the problem is that the tris electrophoresis buffer that the
>491 cell uses has a similar absorbance maximum to our protein (around
>340 nm - collagen has very little UV absorbance at 280 nm).  So, does
>anyone out there know if other (non-tris) buffer systems exist which
>will still give separations around pH 7-8, and which don't absorb so
>strongly at 340 nm ? 
>

Can't help on the buffer but could you not just hook the whole output to
a fraction collector and collect small enough and lots of fractions
based on volume or time i.e. just treat it like column chromatography
Re-analyse the fractions by SDS PAGE to find your protein. More of a
pain with multiple gels to run but also possibly more informative. 

Sorry If I'm repeating what you have already considered and rejected.

Duncan

-- 
The problem with being on the cutting edge is that you occasionally get 
sliced from time to time....

Duncan Clark
DNAmp Ltd.
Tel: +44(0)1252376288
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