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Choosing the best voltage for SDS-PAGE analysis

Choosing the best voltage for SDS-PAGE analysis - SDS-PAGE Gel Electrophoresis Forum

Choosing the best voltage for SDS-PAGE analysis - 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 08-01-2008, 03:47 PM
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Question Choosing the best voltage for SDS-PAGE analysis



Hi everybody

How can we choose the best voltage (current) to run a SDS-PAGE gel?

Best wishes.
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Old 08-02-2008, 05:08 AM
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Default Re: Choosing the best voltage for SDS-PAGE analysis

Hi Bahara,
there have been several papers indicating that constant current is better especially if you use a stacking gel.

If you are only using a resolving gel then I guess the voltage depends on your protein size and gel percentage. For smaller proteins go a bit faster, for larger go a bit slower.
If you are running your gels on large slabs overnite, I have even used 30V.

For regular gels 100V is ok at room temperature with a cold pack, however faster at 110V or 120V in the cold room with a cold pack should be ok.
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Old 08-02-2008, 08:34 AM
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Default Re: Choosing the best voltage for SDS-PAGE analysis

it all depends. most of the time, i just try it,never confirm it.
i think the stacking gel can help a lot to the results.
the voltage between 80-120 is permitable
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Old 08-05-2008, 04:13 PM
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Default Re: Choosing the best voltage for SDS-PAGE analysis

Yes, around 100 V should work quite well. You can also test to see how well your bands resolve either by Coomassie blue staining or Western blotting. If you get sharp bands that are well separated, then that particular voltage should be OK. Try not to run at a too high voltage, as that can heat up your gel too much.
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Old 08-11-2008, 08:45 AM
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Thumbs up Re: Choosing the best voltage for SDS-PAGE analysis

Thank you all guys!
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