gelatin zymography problem - (Nov/07/2007 )

Hi,

I just started running gelatin zymograms and I have a problem that keeps occurring. When I come to destain the gel, the upper half is pale making it difficult to visualise the lytic bands. It seems as if the gelatin is running out of the gel. It is being run at 4 degrees on ice at 10mA per gel. I have tried running at 5 mA per gel as well to see if the resistance is too high so heating up the gel but that made not difference.

Has anyone seen this happen before and know hoe to fix it?

Thanks

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Just wondering...

How do you make the gelatin SDS-PAGE?

Please give more information.

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The lower separating gel (10%) is made up with 3mg/ml gelatin stock and 30% acrylamide.Gelatin stock is heated in water bath to make sure its dissolved.

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