______________________________ Reply Separator _________________________________
Subject: electroelution of proteins
Author: abrf-request@aecom.yu.edu at INTERNET
Date: 05/14/1998 10:15 PM
Hi everyone,
I have questions concerning the electroelution of proteins from acrylamide
gel pieces.
We recently bought a BioRad Model 422 electro-eluter. To start I have just
been trying to elute known quantities of BSA from a 9% mini gel. I then
pass the eluate through a ProBlott cartridge and check the elution amount
and purity through sequencing. So far results have been less than
promising. I have used the standard Tris/glycine/SDS buffer as well as an
NH4HCO3/SDS buffer and elute for 4 hours.
>From sequenicng it appears that very little (<5%) is eluting at this
point.Also the eluate is full of contaminants even after cleanup with the
ProBlott. I am hoping that some of you will have some useful suggestions
and tips to help me improve this technique
Thanks in Advance
Amanda
Amanda Hall
Professional Officer
Newcastle Protein
University of Newcastle
ph 02 4921 7299
fax 02 4921 6903
email: mbah@medicine.newcastle.edu.au