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Too short Resolving Gel and too long stacking gel

Too short Resolving Gel and too long stacking gel - Western Blot Forum

Too short Resolving Gel and too long stacking gel - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 02-24-2011, 07:42 PM
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Default Too short Resolving Gel and too long stacking gel



Normally I like to make my resolving gel go to just under 1 cm below the end of the combs. This time, for whatever reason, I don't know if my resolving gel compressed or what, but it is a full 1.6 cm below the bottom of the combs.

Do you know what sort of problems I can expect from this? Should I expect any problems from this at all?

Thanks!
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Old 02-25-2011, 12:35 PM
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Default Re: Too short Resolving Gel and too long stacking gel

Your gel will still be usable. The major change that you will notice is that your resolving area will be shortened. Meaning the proteins will not have as much distance to separate. So, if you have a complex sample, such as a lysate, you might not get as good of a separation of your bands as you would have with a taller resolving gel.

The major cause of the resolving gel being shorter than you poured is due to leaking during polymerization. This may be due to the casting apparatus or the length of time it takes to polymerize the gel. It is hard to speculate further without knowing your casting protocol.

Hope this helps.
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The Following User Says Thank You to proteaHeather For This Useful Post:
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gel , long , resolving , short , stacking


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