Sephadex LH-20
Sephadex™ LH-20 is prepared by hydroxypropylation of Sephadex G-25, a bead-formed dextran medium, and has been specifically developed for gel filtration of natural products, such as steroids, terpenoids, lipids and low molecular weight peptides, in organic solvents. Characteristics of Sephadex LH-20 are listed in Appendix B, Table 1.
成都朗阁培训中心Instructions 56-1190-97 AD
Gel Filtration
GE Healthcare
Table of contents
1. Preparing the medium suspension 3
2. Packing Sephadex LH-20 5
新生儿护理3. Equilibration 7
4. Eluents 7
英文歌曲大全5. Samples 7
6. Elution 7
7. Regeneration 8
8. Storage 8 Appendix A 8 Appendix B 9 9. Ordering information 10
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1. Preparing the medium suspension
Sephadex LH-20 is supplied as a dry powder and must be swollen before u. During swelling excessive stirring should be avoided as it may break the beads. Do not u magnetic stirrers.
1. Swell the medium for at least 3 hours in an excess of the
solvent to be ud in the paration at room temperature. The extent of swelling will depend upon the solvent system to be ud. The table below should be ud to calculate the amount of dry mediu
m required to give the required bed volume.
2. Prepare a media slurry in a ratio of 75% ttled medium to
25% solvent. Fine particles can be removed by decanting at this time.
Approximate bed volumes (ml media/g dry powder) of Sephadex LH-20 in different organic solvents are given in the table on page 4.
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国家英语四级成绩查询Solvent Approx. bed volume (ml/g dry powder)
Dimethyl sulphoxide 4.4-4.6
Pyridine 4.2-4.4
Water 4.0-4.4 Dimethylformamide 4.0-4.4
Saline 3.8-4.2
Methanol 3.9-4.1coco什么意思
Ethylene dichloride 3.8-4.1 Chloroform1 3.8-4.1
Propanol 3.7-4.0
Ethanol2 3.6-3.9 Isobutanol 3.6-3.9 Formamide 3.6-3.9 Methylene dichloride 3.6-3.9
Butanol 3.5-3.8 Isopropanol 3.3-3.6 Tetrahydrofuran 3.3-3.6
Dioxane 3.2-3.5
Acetone 2.4-2.6 Acetonitrile3 2.2-2.4
Carbon tetrachloride3 1.8-2.2
Benzene3 1.6-2.0
Ethyl acetate3 1.6-1.8
Toluene3 1.5-1.6
1Containing 1% ethanol
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2Containing 1% benzene
3Solvents giving a bed volume of less then about 2.5 ml/g dry media are generally not uful
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2. Packing Sephadex LH-20
cd rom是什么1. Equilibrate all material to room temperature.
2. Resuspend and pour the media slurry into the column in
one continuous motion. Pouring down a glass rod held
against the wall of the column prevents the introduction of
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air bubbles. Fill the rervoir to the top with buffer. Screw on the rervoir top tightly, and connect it to a pump. Open the column outlet.
3. Open the bottom outlet of the column and begin packing at
300 cm/h until the media bed has reached a constant height.
The column should be packed at as high pressure as is
孔雀东南飞剧本possible without deforming the beads (the pressure tolerance of the column is the limity factor, pack at the maximum
pressure specified for your column) Stop the pump, clo
the column outlet and remove the column from the stand.
Unscrew and remove the packing rervoir over a sink.
4. Re-mount the column on the stand and carefully fill with
buffer to form a meniscus at the top of the column.
5. Wet the upper adaptor by drawing solvent through it using
a syringe. Inrt the adaptor at an angle into the column,
ensuring that no air is trapped under the net. Adjust the
adaptor O-ring to give a sliding al on the column wall.南宁会计
6. Make all tubing connections at this stage. There must be a
bubble-free eluent connection between the column and the pump and the column and the sample application system.
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