Bio-Rad Silver Stains - Manual (Page 11)

Section 4
Polyacrylamide Gel Staining Procedure
Before starting this procedure, prepare the working solutions as described on pages 5–8.
Step Description Small Gels Large Gels
(Mini or Criterion (16 × 16 cm
Gels) or larger)
1 Fixing. Place gel(s) in fixing solution >30 min >1 hr
and shake for indicated period of time. (overnight acceptable) (overnight acceptable)
2 Sensitizing. Remove the fixing solution 30 min 30 min
and shake gel(s) in the sensitizing solution.
3 Washing. Remove sensitizing solution 3 × 5 min 3 × 10 min
and wash 3 times with one working
volume of water.
4 Staining. Remove water and shake 20 min 30 min
in staining solution.
5 Rinsing. Remove staining solution 1 × 1 min, (2 × 1 min) † 1 × 1 min, (2 × 1 min) †
and rinse the gel(s) in one working
volume of water.
6 Developing. Remove water and shake gel(s) 10–30 min 10–30 min
in developing solution.
7 Stopping. Remove developing solution and 10 min 10 min
shake gel(s) in stopping solution.
8 Washing. Wash gels in one working volume 10 min 10 min
of water. Gel(s) may be dried, imaged, or used
to excise visualized proteins.
† When using the Dodeca stainer, pay special attention to minimize the solution draining and filling time between steps 5 and 6, and between
steps 6 and 7. However, gels stained individually should be rinsed in step 6 with two 1 min water washes.
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