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
Chemical stability / Any buffer and salt.0.5M NaOH and acetic 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.