Preparative fractionation of amyloid proteins on a microgram scale by high-performance liquid chromatography and polyacrylamide gel electrophoresis.
Preparative separation of amyloid proteins on a microgram scale is presented. Amyloid fibrils solubilized in aqueous 50% acetonitrile containing 0.1% trifluoroacetic acid, are fractionated by reverse-phase high-performance liquid chromatography. Fractionation of amyloids obtained from patients with familial Mediterranean fever allowed isolation of a protein identical with a conventionally isolated AA-protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis is used for preparative separation of AL-proteins. Two protein extraction procedures from Coomassie Blue stained gels are applied using elution in 0.1% sodium dodecyl sulfate containing buffer and 6 mol/l guanidine-HCl solution. The eluted proteins are concentrated and sodium dodecyl sulfate and dye are removed by acetonitrile precipitation of sample.
D000682:Amyloid / Q000302:isolation & purification*; D000686:Amyloidosis / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D010505:Familial Mediterranean Fever / Q000378:metabolism; D006801:Humans; D008832:Microchemistry; D009422:Nervous System Diseases / Q000378:metabolism
D000682:Amyloid