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Polyacrylamide gel electrophoresis

Polyacrylamide gel electrophoresis - Biology Forum

Polyacrylamide gel electrophoresis - Biology Forums. Ask questions and discuss the study of Biology. If you have Biology questions from your homework ask them here!


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Old 11-07-2011, 03:44 PM
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Unhappy Polyacrylamide gel electrophoresis



Hello, I need to analyze a protein solution by SDS-PAGE using a cell with vertical loading. My problem comes right from the beginning, when i fix to the two plates with autoclave tape + parafilm, insert in the form that allows you to load the gel, but often I have overflowing despite this "sealing" tape + parafilm and the silicone rubber sheet on which the support! The screws are tights, so i don't know what to do! if you have tips, please help!
thank you!
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Old 11-24-2011, 10:59 AM
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Default Re: Polyacrylamide gel electrophoresis

In my old lab we use to melt agarose gel. Procedure is take paraffin sheet then pour the melted agarose as straight line and the keep the fixed two plates on the poured agarose. allow the the agarose to solidify, so the bottom of your plates are completely sealed. Now pour your protein gel, there will no leakage.

Hope this tip help you
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Old 11-24-2011, 01:10 PM
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Default Re: Polyacrylamide gel electrophoresis

I assume this means your gel is leaking out of the bottom?

If so, take 1ml of the separating gel (the one you pour first), and add 10ul of TEMED to it. Then pour this down one side and distribute it evenly across the bottom (you may have the vary the volume depending on the thickness/width of your gels - you want a "plug" a couple of mm thick). That volume of TEMED added to that small amount of gel will cause polymerization within 30sec, thus plugging the gel. Same idea as obama's, but without the need for heat.

Bryan
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