Various questions about Silver stain

Dr Engelbert Buxbaum engelbert_buxbaum at hotmail.com
Mon Mar 22 08:27:26 EST 2004


EK wrote:

> I have noticed that it does not matter much if there are air bubbles or not.

Looks ugly and may interfere with quantitation.

> I also don't equilibrate with glycerol. Instead, I use tap water to briefly
> rinse the gel taken directly from the destaining solution, 

The glycerol makes the gel more flexible and aids in rehydration of
cut-out bands, should that become necessary. Literature reports state
that electroelution and reuse of proteins is possible even 10 years (!)
after the gel was archived.

> Yes, I would
> still suggest removing as much as possible air bubbles when covering the gel
> with the second cellophane sheet, but I have not have any cracks even if a
> couple of bubbles still hung aroung the gel. 

If they are _around_ the gel, they are indeed harmless. I just don't
want them _on_ the gel. 

> I also try to do everything
> quick, so that alcohol and acetic acid from the destaining solution don't
> get washed away from the gel. 

The idea with equilibration is to remove in particular the acetic acid
as much as possible to prevent chemical changes in the protein. Remember
that acetic acid is not all that volatile, and rests may stay in the
dried gel.





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