This chapter presents procedures to synthesize polyacrylamide gels containing various copolymerized 6-acrylamidohexyl glycosides in which the sugar is attached to the polymer matrix at the terminal amino group in the aglycon. Glycosides of mono- and disaccharides containing ω-amino groups in the aglycon are prepared. Amino groups are determined using 2,4,6-trinitrobenzenesulfonic acid. Neutral sugars are determined by a phenol-sulfuric acid procedure. The solvent system for thin-layer chromatography is (v/v/v) ethyl acetate-acetic acid-water. The following groups were detected on TLC plates as indicated: sugars, 10% H2SO4 in 50% ethanol; amino groups, ninhydrin spray; acrylamido compounds, ultraviolet absorption or a permanganateperiodate spray. Polyacrylamide gels containing covalently attached carbohydrates have proved to be useful as affinity adsorbents for lectins and in studies with intact eukaryotic cells. Thin, fiat gel pieces have been used as synthetic culture surfaces to show that freshly isolated hepatocytes from chicken or rat will specifically bind to surfaces containing, respectively, N-acetylglucosamine z or galactose.
ASJC Scopus subject areas
- Molecular Biology