SDS-PAGE help badly needed

f.r.byrne gen126 at abdn.ac.uk
Mon Aug 7 08:18:58 EST 1995


sounds like you are doing everything right.
there's always that old reliab;le that
someone borrowed them and neglected
to ask your permission.
make 'em up again yourself and see.
what more can I say?

Fergus
University of Aberdeen


Giorgio Spagnol (spagnol at galactica.it) wrote:
: Dear netters,

: this is really becoming a nightmare! Since two weeks protein samples
: refuse to enter my one dimensional SDS-PAGE minigel.
: Funny thing: only the MW markers enter, as normal.
: Gel is 6% acrylamide, 0.1% SDS, and proteins are of a MW which should
: enter the gel (e.g. 67 kD). I think something is wrong in the loading
: buffer (salts should not interfere, since in the samples concentrations
: are kept in 150 mM or even in H2O  ), but damn if I know what.
: This is the composition of the loading buffer: Tris Cl Ph 6.8,
: Glycerol, 20% dTT O.1M, Blue Bromophenol 0.2%, fial conc. of SDS: 1%. MW
: are in 50% glycerol, 300 mM NaCl, 3mM NaN3, dTT 0.1 M, and I add the
: loading buffer to them as to my protein samples, to keep SDS
: concentration to 1%. Probably all this information is superfluos, since,
: the buffers are the  exactly the same I used before the problem started.
: Please help, or I shall start to believe in black magic, and this is not
: a point of honour for a researcher.
: Thanks!
: Giorgio.



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