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 glycosphingolipi ds from a high-performance thin-layer chromatography (HPTLC) plate to a polyvinylidene difluoride (PVDF) membrane, called thin-layer chromat ography (TLC) blotting, and its application in lipid research are desc ribed. Most of the lipids developed on the HPTLC plate are blotted qua ntitatively. Detection of the lipids on the membrane is done by chemic al and immunological staining. This method makes it possible to purify individual lipid components separated on an HPTLC plate and to determ ine the structures of lipids in limited biological samples when combin ed with direct mass spectrometric analysis (TLC blotting/MS). A bindin g assay and the identification of ligands for microorganisms can be ma de using TLC blotting/MS. In addition, simple methods for the detectio n of enzymes on a substrate-immobilized PVDF membrane are described. ( C) 1997 Academic Press.