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Thin-layer chromatography
of lipids |
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Achieving an optimal
separation of lipids by thin-layer chromatography is not always
an easy task. In addition to the obvious importance of the materials
used (origin of the solvents and silica gel plates), thin-layer
chromatographic separations often greatly depend on environmental
factors such as room temperature and humidity. You were warned! |
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Neutral lipids |
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n-Hexane/ethyl ether/
acetic acid (70:30:1) |
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Phosphatidylethanol
& phosphatidic acid |
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Upper phase of:
ethyl acetate/isooctane/acetic acid/water (130:20:30:100) |
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(Balboa et al.,
J. Biol. Chem. 269: 10511-10516, 1994) |
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Variant one: Upper phase of: ethyl
acetate/isooctane/acetic acid/water (110:50:20:100) |
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(Balboa et al.,
J. Exp. Med. 176: 9-17, 1992) |
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Phosphatidylcholine,
phosphatidylethanolamine, and their lyso forms |
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Chloroform/methanol/acetic acid/water
(50:40:6:0.6) |
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(Balsinde et al.,
J. Biol. Chem. 272: 29317-29321, 1997) |
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Variant one (less
acetic acid): chloroform/methanol/acetic
acid/water (65:50:1:4) |
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Variant two (more
acetic acid): chloroform/methanol/acetic
acid/water (50:25:8:4) |
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Phosphatidylinositol
and lysophosphatidylinositol |
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Chloroform/methanol/water/ammonia
(45:35:8:2) (Plates
sprayed with 1% potassium oxalate should be used) |
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(Balsinde et al.,
Biochim. Biophys. Acta 1136: 75-82, 1992) |
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Phosphatidic
acid and lysophosphatidic acid |
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Chloroform/methanol/10
M HCl (87:13:0.5) (Plates sprayed with 1% potassium oxalate should
be used) |
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(Billah et al.,
J. Biol. Chem. 256: 5399-5403, 1981) |
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Two-dimensional
separation of major phospholipids |
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System I: chloroform/methanol/water/11M
NH3 (65:25:1.75:2.5) |
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System II: chloroform/acetone/methanol/acetic
acid/water (30:40:10:10:5) |
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to Protocols |
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