Bio-Rad Aurum Ion Exchange Mini Kits - Manual (Page 14)

12
Section 7
Troubleshooting Guide
Problem Possible Cause Possible Solution
Low protein Low levels of protein Increase load on column
concentration in sample
Concentrate sample in
SpeedVac
High protein High levels of protein Decrease the load
concentration in sample applied to column
Buffer does not flow Frit is clogged with Prefilter sample prior
through column sample debris to loading
Protein does not bind Incorrect column chosen CEX column is for
to column basic proteins; AEX
column is for acidic
proteins
Ionic detergents can Avoid using detergents
prevent sample from with opposing net
binding charges to that of ion
exchange column.
Detergents can be
removed using
Bio-Beads
® SM-2
macroporous beads
(catalog #152-3920)
Ionic strength too high Dilute sample or desalt
pH not correct Check pH of loading
buffer
Protein does not elute Salt concentration too Increase salt
from column low in elution buffer concentration to 2 M
and/or change pH
Sample is dilute after Excess packing buffer Remove excess packing
centrifugation was not removed buffer by centrifugation
prior to loading sample
4110137B-4.0.GR.qxd 8/14/2003 2:24 PM Page 12