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IEF problem: the tracking dye did not migrate during the run.

IEF problem: the tracking dye did not migrate during the run. - Proteomics Forum

IEF problem: the tracking dye did not migrate during the run. - Post Questions and Discuss Proteomics, Proteomic Bioinformatics, Proteomic Techniques such as 2-D, Mass Spec etc.


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Old 10-10-2007, 03:39 AM
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Default IEF problem: the tracking dye did not migrate during the run.



I'm new in this field. I tried to run my sample in the 13 cm strip (Amersham). After overnight rehydration, I programmed the IEF to start automatically. I don't know why the tracking dye did not migrate to the anode. What's the problem? the strip holder or the rehydrated strip? Did anyone have the same problem? Please tell me how to figure out.

Thanks in advance
ay_sh
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Old 10-10-2007, 04:02 PM
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Default Re: IEF problem: the tracking dye did not migrate during the run.

1. Do you have ampholytes in rehydration buffer?
2. Usually you have a stable resistance when isoelectric separation is complete, so you can monitor that.
3. Electrode wicks are very beneficial, use them. That means adding them in between ON rehydration and beginning the IEF run.
4. Make sure you aren't using too much protein/too much dye.
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