Fw: Troubleshooting IEF
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
I can't be a lot of help - we were doing 2D electrophoresis and our IEF was
done on strip gels.
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
> 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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