SDS-Agarose electrophoresis of HMW proteins

[David Micklem]

Hi,

Someone here mentioned recently the use of SDS-Agarose gel
electrophoresis for separation of large proteins (I am interested in
>>500kDa proteins/complexes).

Can anyone point me to a good practical protocol for this method? Its
not in any of the general lab manuals that I've been able to lay my
hands on. Any comments on general limitations of the method compared
to SDS-PAGE?

I am particularly interested to know if it is possible to run
'diagonal' 2D gels (non-reducing 1st dimension, reducing 2nd
dimension) in SDS-agarose. Can the gels be stained/blotted/etc the
same way that SDS-PAGE gels are? What is the approximate range of
sizes that can be separated on a single gel - presumably that is
dependent on the agarose concentration?

AdvTHANKSance,

David

-- 
D.R. Micklem,              Time flies like an arrow... 
Dept. of Genetics,          Fruit flies like a banana.
Cambridge University,      Email:drm21 at mole dot bio dot cam dot ac
dot uk.
Cambridge, UK              Phone: +44 (1223) 766336
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