Isoelectric Focusing

Darren Tyson tysondr.nospam at slu.edu
Thu Nov 6 14:55:56 EST 1997



D. Vadnais wrote:

> I am going to use IEF slab gels from Bio-Rad to separate invertase
> isozymes from alfalfa.  I have never done this procedure before, and I am
> not really sure of the protocol.  I have only the protocols that are used
> for 2D gel electrophoresis using the capillary tubes.  I need to know
> what buffers I should be using to run these gels, and what type of
> loading buffer I require for my samples.
> Thanks in advance for any information you can give me.
> Dave

Dave,

I use the BioRad IEF precast gels which have well in my hands.  The cathode
buffer is 20 mM lysine, 20 mM arginine.  The anode buffer is 7 mM phosphoric
acid.  Running conditions are 100 V for 1 hr, 250 V for 1 hr and 500 V for 30
min.

Hope this helps.

Darren





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