Instruction
DEAE-Cellulose, the mean particle size of which is 100μm, are hydrophilic high polymers.
The surface of the polymers is grafted with sugar macromolecules. So it has higher
specific surface area and better bio-compatibility and maintains a high capacity under high
flow rate. At the same time, the resolution is very good. Benefit from the high specific
surface area, the equilibration and elution processes are very fast. The capacity remains
unchanged when it is even used to purify super large molecular such as virus or plasmids
after grafting.
This product has good physical and chemical stability, so it can be used in long term and
is very convenient to use with stable capacity
Gel characteristics
Feature
High capacity, good resolution, high flow
rate, convenient to use
Bead structure
Highly cross-linked cellulose
Ligand
Diethyl amino ethyl
Ligand density
40μmol /ml
Absorption capacity
150mg HSA/ml
Particle size
50-250μm
Max. flow rate
200cm/h
pH working range
3-10，onsite cleaning:2-11
Any buffer and salt.0.5M NaOH and acetic
Chemical stability
acid,8M urea，6Mguanidine hydrochloride,
ethanol, isopropanol and so on.
Physical stability
0.1M neutral buffer，120℃30min
Storage temperature
+4~30℃
Storage
Dry powder，1g=3-4ml
Application notes:
（1）Packing the column
1．Equilibrate all material to the temperature at which the chromatography will be
performed. The liquids should be degassed. The required amount CM-Cellulose can be
directly weighted and swelled in the buffer for 1 hour (it is better to soak the cellulose in
appropriate amount double-distilled water for 12hours before swelling with buffer)
2．Add 20% ethanol to the bottom of the column to remove the air, then close the column
outlet and keep a little 20% ethanol in the column. The 20% ethanol is prone to bubbles
where 1% Tween is added to avoid bubbles. The ethanol can be replaced with pure water
if you want to pack the column with pure water. In this case, it needs a suction funnel. You
should carefully decant the 20% ethanol and replace it with 5 volumes pure water and
repeat precipitate, then pack the cellulose after five times.
3．The cellulose particles are relative small, so make sure the screen mesh is right to
eliminate leakage. You can take a try by adding a few particles on the screen mesh to
make sure no problem. Then pour the slurry into the column in one continuous motion.
Pouring the slurry down a glass rod held against the wall of the column will minimize the
introduction of air bubbles. Natural settlement until the volume does no longer change. At
this time, the cellulose and the liquid delaminate well. The upper solution is clear. Then,
the pump can be open at an appropriate flow rate to press the column until the volume of
the cellulose does no longer change, then tight the convert to the cellulose closely. The
column can be used after equilibrating under the flow rate when packing.
4. Place the cellulose at room temperature for 2-3hours after unpacking from the
refrigerator and before packing the column to avoid bubbles resulting from the
temperature changes.
（2） Binding
The salt concentration and pH of the samples should be the same as the balance buffer.
The protein can’t bind if the salt concentration is too high or the pH is too low. Make
adjustment according to the samples.
（3） Elution
If elute with linear gradient, the radio of the column height/diameter should be greater than
10, the greater the radio, the better the resolution. It is better to load small amount
samples. The sample amount applied should be approximate 10mg/ml. If elute with step
gradient, short and thick column can be used, more important is that there is no limit on
the sample volume. It is easy to amplification. The repeatability is good. If the elution
condition is good, the results can be as good as the linear gradient or even better.
Choose the method according to what you need. Regeneration and cleaning
1．Wash the column with 5 bed volumes buffer (0.5MNaOH, 2M NaCl), and then 5 bed
volumes water every time. The medium should be store in 20% ethanol. After 3-5times,
the medium should be washed with 5 bed volumes 70% ethanol or 30% isopropanol, both
of which contain 1% Tween, after washing with water. Wash the column with 5 bed
volumes 20% ethanol at last.
2．The mixture of organic solvent and water is prone to bubbles, so the prepared organic
solvent should be stored at room temperature overnight to avoid introducing air bubbles
so that the column is not working.