Bio-Rad Bio-Rex® Reactor Grade Resins User Manual
Page 7
Fig. 2. Separation of hemoglobin from ampholytes
and sucrose on a mixed bed ion exchange column
(AG 501-X8 resin).
A hemoglobin blank in the sucrose
determination gives an absorbance of about 0.1. Neither
sucrose nor ampholyte emerged from the column in the
total volume tested.
3
AG 501-X8 resin has also been shown to be useful
for separating peptides of greater than 4,000 daltons from
ampholytes.
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Table 3. Deionization of Urea by Batch and
Column Methods
Batch
Column
Sample
100 ml 6 M urea
100 ml 6 M urea
Starting conductivity
70 µmho/cm
70 µmho/cm
Amount of mixed bed resin
5 grams
5 grams (~8 ml)
Final conductivity
5.0 µmho/cm
0.2 µmho/cm
Time
~5 hours
~10 minutes
Ampholyte Removal
Carrier ampholytes may be quantitatively removed
from protein fractions derived from isoelectric focusing
using mixed bed resin. Mixed bed ion exchange chro-
matography represents a method for the quantitative
removal of carrier ampholytes. Figure 2 illustrates the
separation of proteins from ampholytes and sucrose. A 3
ml sample was applied to a 0.9 x 25 cm column of AG
501-X8 resin. AG 501-X8 resin has also been shown to be
useful for separating peptides of greater than 4,000 dal-
tons from ampholytes.
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LIT205B 6/17/98 12:15 PM Page 10