reversible stain? - (Nov/10/2006 )

Hello:

Do I need to destain a sypro ruby stained gel?I´d like to stain again with a silver protocol.

Thank you!

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Hi bertis,
here you are link that may be useful to figure out the difference between staining/destaining/restaining and sequential staining. You'll decide later on which one better suits your needs
http://aiche.confex.com/aiche/2005/techprogram/P23050.HTM

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I think that sypro cannot be destained, at least I have never seen a protocol about it and never heard it.

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At least it can be destained from plugs Sypro Ruby stained, as follows
Staining Gels with Sypro Ruby and Destaining Gel Plugs
1. Rinse gel with deionized water.
2. Incubate gel in fixative (30 % ethanol / 7.5 % acetic acid) for at least two hours.
3. Rinse gel with deionized water.
4. Place gel in polypropylene, polycarbonate, or polyvinyl chloride tray and cover with
Sypro Ruby stain.
5. Incubate gel for five hours to overnight with gentle shaking, protected from the light.
6. Rinse gel with deionized water.
7. Incubate gel in wash (10 % methanol / 6 % acetic acid) for 1-2 hours.
8. Working in a dust- free environment, image gel (use 400 nm excitation / 633 nm
emission, but Sypro Ruby can be imaged using standard UV transillumination.) and pick
spots. Place gel plugs in clear polypropylene tubes/plates.
9. Incubate gel plugs with 100 ml 50 mM ammonium bicarbonate in 50 % methanol for 20
minutes.
10. Remove methanol solution. Repeat.
11. Incubate gel plugs with 100 ml 75 % acetonitrile for 20 minutes.
12. Remove acetonitrile solution. Dry gel plugs at 40 oC for 10 minutes.

You may try to perform steps 9-12 adjusting times

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Wow, very interesting!! Thanks lillymay

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