Bio-Rad Negative Stains - Manual (Page 6)

Section 3
Additional Information
3.1 Commonly Asked Questions
1. Can I blot the copper Yes, after destaining. The Copper
stained gel? Stain reversibly fixes the proteins in
the gel.
2. Am I able to stain the gel Yes. In fact, only with a silver stain
later with either CBB must you first destain the gel. The
or silver? low pH of the Coomassie staining
solution acts as a destainer.
3. Can the Copper Stain be For optimum results, discard the
reused? solution after each use. Results show
that it might work a second time, but
with each use the staining is less
effective.
4. Can the Copper Destain be No, this is not recommended.
reused?
5. Can I stain DNA in a Bands do not develop when copper
TBE gel? staining a DNA gel.
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3. Replace this rinse with fresh, diluted Copper Destain and gently
agitate again for the recommended time. Repeat with 1:20 Copper
Destain, if recommended in Table 2.
4. The gel is now ready for Coomassie staining, silver staining,
blotting, or other analyses.
Table 2. Copper Destaining Wash Times
The following are recommended destaining wash times. The
destaining procedure has been optimized for mini-gels. Slight
variations in the protocol may be necessary to optimize results with
full-size gels.
Gel Thickness Destain #1 Destain #2 Destain #3
0.5 mm 5 minutes 3 minutes ——
0.75 mm 5 minutes 5 minutes 5 minutes
1.0 mm 5 minutes 10 minutes 5 minutes
The Copper Destain solution is diluted 1:10 with water. If necessary,
Destain #3 can be further diluted to 1:20, which will help to conserve
reagents.
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