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