title: Optimal resolution of eye lens gamma-crystallins by cation-exchange high-performance liquid chromatography on SynChropak CM300.
abstract: Cation-exchange high-performance liquid chromatography on SynChropak CM300 in Tris-acetate buffers of pH 5-7, using sodium acetate gradients, produces an excellent separation of the various gamma-crystallin gene products and their post-synthetically modified forms from eye lens. With a single analysis of total lens extract, the gamma-crystallins can be resolved, quantified and collected for amino acid analysis. Experimental conditions are presented for optimal resolution of individual human, rat, bovine and dogfish shark gamma-crystallins. Applications presented include determinations of different synthesis of gamma-crystallins and chemical modification (oxidation by hydrogen peroxide) in situ.
authors.affiliation: Department of Physics, Massachusetts Institute of Technology, Cambridge 02139.
authors.forename: R J
authors.initials: RJ
authors.lastname: Siezen
authors.forename: E D
authors.initials: ED
authors.lastname: Kaplan
mesh_terms: D000375:Aging / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003459:Crystallins / Q000032:analysis*; D004284:Dogfish; D006801:Humans; D007223:Infant; D007475:Ion Exchange Resins; D007908:Lens, Crystalline / Q000032:analysis*; D051381:Rats; D013045:Species Specificity
chemical_list: D000596:Amino Acids; D003459:Crystallins; D007475:Ion Exchange Resins