Silver Staining or SYPRO Red
AndrewLiddell at nospam.Hotmail.com
Wed Aug 18 20:47:24 EST 1999
Try adding 0.1 % w/v sodium thiosulphate to your gel mix (added from a 5%
w/v stock kept at 4oC, renewed weekly). If you do this you will also have to
double the amount of TEMED and ammonium persulphate used to achieve
polymerisation in a similar time. This seems to have a significant effect in
reducing silver stain backgrounds and no effect on resolution/run times etc.
In addition, or alternatively, you could try using a different crosslinker
such as piperazine diacrylamide (PDA) in place of the usual 'bis'. PDA is
available from Bio-Rad and can also reduce background stain but is a bit
more expensive than 'bis'.
One other thing which may be of interest - Molecular Probes now supply Sypro
Ruby for protein gel staining which is very expensive but they report that
it is very sensitive, I haven't any experience of using it yet though.
I don't have the reference for the thiosulphate technique to hand but if you
are interested contact me at Andrew.Liddell at plants.ox.ac.uk and I will find
it for you.
AO <andrewo at bigfoot.com> wrote in message
news:379fb6aa.5037874 at news.supernews.com...
> Does anyone have any suggestions on reducing the amount of
> background when silver staining. Also if anyone has any modifications
> to the procedure that speeds it I would be eternally grateful. On
> another note, has anybody used SYPRO Red or Orange. How well do they
> Andrew Osterburg
> osterbar at email.uc.edu
More information about the Methods