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Staining 2D Gels

Staining 2D Gels - 2-D Gel Electrophoresis Forum

Staining 2D Gels - 2-D Gel Electrophoresis Forum. A forum dedicated to two dimensional gel electrophoresis, discussion about systems, and troubleshooting its problems.


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Old 07-01-2009, 02:24 AM
aoi aoi is offline
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Question Staining 2D Gels



hello all.

i used silver staining for 2D gels but somehow my gels turn out to be very dark (making the spots hard to see). can anyone gives opinion/suggestion?

isn't necessary to wash the gel with ethanol after fixation (before sensitizing)?
and im thinking of reducing the time for sensitizing and developing steps.
ty
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Old 02-03-2010, 05:37 PM
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Default Re: Staining 2D Gels

Look up Invitrogen's silverquest protocol. All steps are 10 min except staining (15 min) and developing (~15 min).

You should wash after fixing, sensitizing, and even staining (stain wash is with water for 60 sec)
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Old 03-17-2011, 06:50 PM
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Cool Re: Staining 2D Gels

Hi aoi,
The problem of darkening comes if you develop the gel for longer times.....keep the stop solution ready by ur sides the moment u start developing the gel.....transfer the gel to stop solution after adequate developing......

hope this helps....
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Old 03-17-2011, 06:53 PM
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Default Re: Staining 2D Gels

Hi,
can anyone tell me how to avoid or to clean the light brownish background in my silver stained gels.......
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Old 10-28-2011, 10:11 AM
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Default Re: Staining 2D Gels

Hi

High sensitivity identification using LC-ESI-MS/MS (as opposed to lower sensitivity MALDI-TOF/TOF-MS) can cost $200 per spot or more.
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