I am not using an IPGphor. However, using a Multiphor at 5000 V, I have usually
around 20-40 microamp/strip at steady state. Either you have not desalted your
sample properly or you have way too much ampholines in your sample buffer
(around 0.5%-1% is usually the rule).I do not believe that the separation will
be to your expectation with the amount of current you mentioned in your message.
Hope that this helps.
Axel
jaime paba martinez wrote:
> I am testing the IPGphor system from Pharmacia. The users manual suggests
> a 50microAmp/strip during focusing, but it does not allow the voltage to
> raise until 8000volts in the four hours protocol. I would like to know if
> someone has used a current higher than 200microamp/strip during focusing
> with a good result.
>
> thanks in advance
>
> Jaime Paba
> Biochemistry Lab
> University of Brasilia
------------------------------------------------------------------------------
--Axel Ducret, Ph.D. Senior Research Biologist Merck-Frosst Canada Inc. Dept. Biochemistry and Molecular Biology P.O. Box 1005 Pointe-Claire-Dorval PQ H9R 4P8 Canada
tel. + (514) 428-3428 fax + (514) 428-4900