megadalton protein purification

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Wed May 31 09:24:48 EST 2000


In article <3934711E.B33DB9C8 at ppserver.tamu.edu>, Aric Wiest <
awiest at ppserver.tamu.edu> wrote:
>Hi,
>
>Does anyone have any ideas on how to isolate a protein that may
be as
>large as 2.4MD.
>
>Thanks,
>Aric Wiest
>

In addition to "normal" chromatographic steps (like ion-exchange,
etc.,) the following might be useful:

- Sepharose 2B or 4B

- Velocity sedimentation of sucrose or glycerol gradients (Off
the
top of my head, it seems like that might be about as big as a
ribosome subunit, so in the 40S-60S range)

- PEG precipitation

Heck, you could probably pellet it at 100000 x g

Is this beast a single chain (!)? A complex? Soluble? Membrane-
bound?


Nick Theodorakis

nicholas_theodorakis at urmc.rochester.edu
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