2D nightmare

David C. Logan david.logan at plant-sciences.oxford.ac.uk
Sun Jun 9 07:18:52 EST 1996


>(deleted) Has anyone ever gotten this thing 
>to work?  All advice, specific and general, is welcome.                        
>         
>--jgluz at stud.med.cornell.edu  


>Hi,
>Supposedly, if every condition you have used is right, you should  
>observe a decrease in the current conductivity after the proteins have  
>reached their pIs. At this point, the proteins can not conduct the  
>current.
>Therefore you encountered a very high resistance, according to V=IR,  
>where V is voltage, I is current and R is resistance.  You should set  
>constant amperage to run the IEF gel, but not constant voltage.   

>best wishes,

>Chi-Kuang Wen

Well, the original post did state that migration stopped due to too low 
a current after 30 mins - this is a fast IEF run if already at 
equilibrium. Secondly, the ampholines or immobilines in the system would 
be unlikely to be focused (ie. at their pI) after such a short run 
(assuming no prefocusing stage which is an unecessary step with this 
system). Third, the run should be constant voltage since the current 
will decrease during the run and if left constant at a high level could 
result in too much heat. 
***************************
david.logan at plants.ox.ac.uk

David C. Logan
Department of Plant Science
University of Oxford
South Parks Road
Oxford
OX1 3RB

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