Dear David,
I use colloidal coomassie all the time:
17% w/v ammonium sulphate
3% v/v ortho-phosphoric acid.
34% methanol
0.1% w/v G-250
Add the ammonium sulphate, methanol and phosphoric acid to the water. It
may take some time to dissolve the ammonium sulphate (try heating and
stirring or sonicating), but it is important to dissolve all of the
reagents *before* adding the G-250.
The stain will bind preferentially to the proteins, not the gel, so there
is limited background staining. I use 1% acetic acid to remove what
background staining is present though. For best results, leave the gel in
stain for up to 3 days. It looks even better (fainter spots/bands
enhanced) if you stain-destain-stain again. But this also increases
methylation of the proteins.
Let me know if you require other details.
Regards,
Amanda Nouwens.
>I was wondering if anybody could help me with a "recipe" for
>making up Collodial Blue (Comassie G-250) solution for
>staining SDS-PAGE gels. Do you just make it up in water?,
>and if so what sort of concentration?
>>Any help would be appreciated
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Amanda Nouwens
Australian Proteome Analysis Facility (APAF)
Level 4, Building F7B
Macquarie University
Sydney, NSW 2109
AUSTRALIA
Tel: 61 2 9850 6209 (office)
Tel: 61 2 9850 6255 (lab)
Mob: 0417 237 457
Fax: 61 2 9850 6200
http://www.proteome.org.au
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