help on silver staining PAGE needed......

[BS04%UK.AC.DUNDEE.PRIMEB at CORNELLC.CIT.CORNELL.EDU]

I hope some one can save me reinventing the "wheel". I have recently carried
out some native PAGE 7.5% on some crude extracts of Phaeodactylum tricornutum
a marine diatom. The crude extracts have low protein levels about 30 ug /
100 uL and staining with CBB just shows 4 bands at the most. The most
abundant protein is RUBISCO the main carboxylase. I have thought about silver
staining and this is where I need the advice/help.

A morning in the library and I came up with Switizer, Merril and Shifren
(1979) and Oakley, Kirsh and Morris (1980). The latter appears to be a good
modification of the former but reading more recent papers it does not seem
to get quoted all that often. We have a Phastsystem for electrophoresis
but I would like to some 20x20 cm gels ago and stain with Silver.


I would like to know if anyone can recommend the current method for silver
staining polyacrylamide gels.

Many thanks.........

Andy J.






Andrew M. Johnston,                    E-Mail: BS04 at PRIMEB.DUNDEE.AC.UK
Biological Sciences,                   Tel No: 0382-23181 Ext 4282
University of Dundee,
Dundee
Scotland
DD1 4HN                                DUNDEE UNITED GIVE A CHAP A HARD TIME!!!