title: TLC-Blot (Far-Eastern Blot) and Its Application to Functional Lipidomics.
abstract: A simple method for transfer of lipids-including phospholipids, glycolipids, and neutral lipids-from a high performance thin layer chromatography (HPTLC) plate to a polyvinylidene difluoride (PVDF) membrane, TLC-Blot (Far-Eastern Blot), and its biochemical applications are presented. This chapter presents the conventional procedures for separating lipid from tissue samples, cultured cells, and serum and the subsequent development of TLC. Individual lipids separated on an HPTLC plate can be transferred to the PVDF membrane quantitatively and also isolated from the lipid-blotted membrane by a one-step purification procedure. Immunodetection with monoclonal antibodies and treatment with lipid-metabolizing enzymes on the lipid-blotted membrane are possible. The method for identification of individual lipids transferred on the PVDF membrane using matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (TLC-Blot/MALDI-TOF MS) is shown as a functional lipidomics application.
authors.affiliation: Niigata University of Pharmacy and Applied Life Sciences, 8-4 Sanoumiya Ejiri Kitajima-cho, Tokushima, 771-0205, Japan, t.taki1212@athena.ocn.ne.j.
authors.forename: Takao
authors.initials: T
authors.lastname: Taki
mesh_terms: D000818:Animals; D002855:Chromatography, Thin Layer / Q000379:methods*; D019295:Computational Biology / Q000379:methods; D005732:Gangliosides / Q000032:analysis; D006017:Glycolipids / Q000032:analysis; D006028:Glycosphingolipids / Q000032:analysis; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D008055:Lipids / Q000032:analysis*; D008567:Membranes, Artificial; D051379:Mice; D010743:Phospholipids / Q000032:analysis; D011145:Polyvinyls / Q000737:chemistry*; D051381:Rats; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D005465:Fluorocarbon Polymers
chemical_list: D005732:Gangliosides; D006017:Glycolipids; D006028:Glycosphingolipids; D008055:Lipids; D008567:Membranes, Artificial; D010743:Phospholipids; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers