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Simultaneous Separation and Purification of Neurofilament and Glial Filament Proteins from Brain
Author(s) -
Liem Ronald
Publication year - 1982
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1982.tb10865.x
Subject(s) - neurofilament , protein filament , urea , ionic strength , hydroxylapatite , chromatography , chemistry , biophysics , intermediate filament , gel electrophoresis , electrophoresis , biochemistry , cytoskeleton , biology , organic chemistry , immunology , immunohistochemistry , aqueous solution , cell , enzyme
Neurofilaments (NF) and glial filaments (GF) were purified from bovine brain by the axonal flotation method, followed by hydroxyl‐apatite chromatography in 8 m ‐urea. The proteins were shown to be competent to reassemble into intermediate filaments with removal of the denaturant, and reassembly was used as the final step in the purification of the filament proteins. The reassembly was found to be dependent on ionic strength and pH. This dependence was greater for neurofilaments than for the glial filaments. The NF and GF preparations were found not to be contaminated with each other by their gel electrophoretic profile and their immunological distinctness. The filament proteins can be obtained in high yield, and remain in solution if the urea is removed by dialysis against a low‐ionic‐strength buffer. Hence, they can provide a source for further biochemical studies.