Fw: Troubleshooting IEF

Deanne Bell dbell at qnis.net
Fri Jan 5 11:46:55 EST 2001


Have you quantified your protein with a Bradford or Pierce's protein assay?
Do you know how much protein you are loading?
What electrophoresis system are you using?
I recommended the electrophoresis discussion group @ Amersham Pharmacia's
web site.
I can't be a lot of help - we were doing 2D electrophoresis and our IEF was
done on strip gels.

Good luck
Deanne Bell
Molecular Markers Lab Technician
USDA Agricultural Research Service
2021 S. Peach Ave.
Fresno, CA 93727
e-mail: dbell at qnis.net
phone: 559 453-3170
fax: 559 453-3088

----------
> From: Kvita <kvita at yahoo.com>
> To: methods at hgmp.mrc.ac.uk
> Subject: Troubleshooting IEF
> Date: Thursday, January 04, 2001 4:11 PM
> 
> Hello,
> My name is Olesia. I am an undergraduate student
> attempting to characterize an experimental protein
> with IEF. Unfortunately, the first 7 gels showed
> nothing: we were able to see standards somewhat
> clearly, but none of the sample proteins showed. We
> tried changing conditions, such as duration of the
> run, making new batches of the cathode and anode
> buffers and staining solutions, varying
> voltage/current, etc. Can you please give me some
> ideas what may be causing this problem. I would also
> appreciate if you could direct me to any good internet
> resources on IEF.
> Thank you very much,
> you can contact me by e-mail: kvita at yahoo.com
> 
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