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A proper ratio of stacking & separating gel

A proper ratio of stacking & separating gel - SDS-PAGE Gel Electrophoresis Forum

A proper ratio of stacking & separating gel - SDS-PAGE Forum and Gel Electrophoresis Forum. Discuss the running of agarose gels, sds-page gels, and other gels including sequencing, gradient or tricine.


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Old 04-25-2008, 08:12 PM
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Default A proper ratio of stacking & separating gel



Hello everybody,
My problem is that I have a large volume of a protein sample. But my capacity of the gel pockets are restricted. Now my question can I pour a gel with a long stacking gel that my volume will fit properly? Because I don`t want to precipitate my samples. Or will I get problems? I am not sure. Once I read one should have a long separating gel. My mw of my protein is about 180kDa.
Please help me.
Thanks
Tiffy
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Old 06-23-2008, 05:53 AM
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Default Re: A proper ratio of stacking & separating gel

Hello,
i sometimes get by this by using larger (thicker) combs which allow about 50ul to be loaded however depending on the amount you need to load and your gel you may need as you say to increase your stacking gel.

If your lab or another lab next to you has access to large (and i mean large) slab gels, I used to run these with up to 200ul in the lanes. These are huge slab gels for autoradiography and can be found in many labs i would just borrow them but ask if they have been used with radioactivity as you may want to clean them before you use them.

cheers
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