Pharmacia Biotech Multiphor II User Manual page 27

5. Operation
24
Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com
Detection
For automated silver and coomassie staining of polyacrylamaida gels see
Protocol guide, Hoefer Automated Gel Stainer (80-6343-34).
Coomassie staining
On completion of the electrophoresis, immediately immerse the gel in
Staining Tray 1 using the solutions and times indicated in table below.
The staining and destaining steps should be carried out on a shaking table.
(See section 4.9 for stock solutions). Each step requires 250 ml of solution.
Step No Solution
1 Fixing solution C
2 Destaining solution I
3 Staining solution K
4 Destaining solution I
5 Destaining solution I
6 Preserving solution L
The staining solution should be heated to 60 °C and poured over the gel.
No further heating is necessary. Destain the gel using several changes of
destaining solution (I) until the background is clear. Change the solution
frequently (particularly at the beginning) in order to speed up the
destaining. To preserve the gel, soak a cellophane sheet in preserving
solution (L). Place it on the gel surface. Remove any air bubbles and wrap
the excess cellophane around the glass plate. An additional glass plate may
be placed underneath during drying to stop the cellophane from shifting or
wrinkling. Leave the gel at room temperature until it is completely dry.
Silver staining
Silver staining is performed essentially as described by. J. Heukeshoven and
R. Dernick, Electrophoreses Forum 1986, 22-27.
On completion of the electrophoresis, immediately immerse the gel in
Staining Tray 1 using the solutions and times indicated in the table below.
All steps should be carried out at room temperature in daylight, while
gently shaking the solution. Use 250 ml of solution for each step.
(See section 4.9 for stock solutions).
Time (min) Temp
°C
20 23
2 23
10 60
20 23
30 23
10 23