Loading capacity of PAGE

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Thu Jan 6 14:01:36 EST 2000


In article <3874CA90.B23DF124 at cellbio.unige.ch>, Bruno Cenni
<cenni at cellbio.unige.ch> wrote:
> Hi all,
> I was not able to find it in my books, so I'll try it here:
> what is the maximal protein loading capacity of standard SDS-PAGE
> gels
> for which you still get resolved bands?
> eg. a 0.75 mm thick "mini"gel (1 lane maybe 5mm wide) with 7-15%
> vs a
> 1.5 mm "midi" gel (lanes about 10 mm)
> TIA
> Bruno


When I used to do a lot of in vitro translations with retic. lysate, I
would load a couple of hundred micrograms of protein on a single lane
on a 1.5 mm gel. Of course, >90 % of it was globin, but the other bands
would resolve fine. Make sure you have enough SDS to bind all the
protein in the sample.

Nick



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