TLC blotting: application to microscale analysis of lipids and as a new approach to lipid-protein interaction.
A simple method for the transfer of phospholipids and glycosphingolipids from a high-performance thin-layer chromatography (HPTLC) plate to a polyvinylidene difluoride (PVDF) membrane, called thin-layer chromatography (TLC) blotting, and its application in lipid research are described. Most of the lipids developed on the HPTLC plate are blotted quantitatively. Detection of the lipids on the membrane is done by chemical and immunological staining. This method makes it possible to purify individual lipid components separated on an HPTLC plate and to determine the structures of lipids in limited biological samples when combined with direct mass spectrometric analysis (TLC blotting/MS). A binding assay and the identification of ligands for microorganisms can be made using TLC blotting/MS. In addition, simple methods for the detection of enzymes on a substrate-immobilized PVDF membrane are described.
Taki
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Faculty of Medicine, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo, 113, Japan. t_taki@research.otsuka.co.jp
Ishikawa
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D000818:Animals; D002855:Chromatography, Thin Layer / Q000379:methods*; D006028:Glycosphingolipids / Q000737:chemistry; D006801:Humans; D008024:Ligands; D008055:Lipids / Q000737:chemistry*; D008567:Membranes, Artificial; D008832:Microchemistry; D011145:Polyvinyls; D011506:Proteins / Q000737:chemistry*; D005465:Fluorocarbon Polymers
D006028:Glycosphingolipids; D008024:Ligands; D008055:Lipids; D008567:Membranes, Artificial; D011145:Polyvinyls; D011506:Proteins; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers