Iso-Electric Focusing Questions
joan at med.unc.edu
Thu Apr 22 13:21:58 EST 1993
I'm having problems with my IEF!!
Let's see: I've been pouring my own gels and they seem to be pouring fine
but when I run them I only get staining (and not very good) on one end
(right-hand side) of the gel. I'm using a flatbed set-up, power: 4 watts,
9 (starts at 9 and goes down to about 3.5) miliamps, and about 1000 volts
(starts at about 400 and goes to 1000). I'm using a mix of ampholytes:
5.0-7.0 and 3.5-10.0 from Pharmacia.
1. Would a too low voltage cause the samples to run only on one side of
2. Would a precast gel (5.0-6.5) be best?
3. How about pre-blended ampholytes? (Pharmacia makes a 5.0-6.5
4. How hard is it to successfully pour an IEF gel? One gel I did worked
but I haven't had any luck since.
5. Anything I should look at that I haven't already looked at? (I've
checked the stain and it's ok, I've pour straight SDS/PAGE and gradient
gels that worked)
7. e-mail would probably be best
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