Optimal resolution of eye lens γ-crystallins by cation-exchange high-performance liquid chromatography on synchropak CM300

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 γ-crystallin gene products and their post-synthetically modified forms from eye lens. With a single analysis of total lens extract, the γ-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 γ-crystallins. Applications presented include determinations of differntial synthesis of γ-crystallins and chemical modification (oxidation by hydrogen peroxide) in situ.