Purification of glial fibrillary acidic protein (GFAP) from normal bovine brain.
Glial fibrillary acidic protein (GFAP) was purified from normal bovine brain by a modification of the procedure used to isolate vimentin in order to avoid contamination by other cytoskeletal components: vimentin, neurofilament triplet proteins, tubulin and actin. GFAP is thought to be separated from vimentin in the DE cellulose column chromatography step. The three other major proteins were also separable through ion exchange and gel filtration column chromatographies. A purified 49 kDa polypeptide was estimated to be GFAP from peptide mapping and subsequent immunoblotting analysis. We obtained 4.4 mg GFAP/1 g bovine brain white matter in less than 3 days. The polyclonal antibody raised against purified GFAP was able to detect 49 kDa GFAP by immunoblotting analysis. This isolation method is simpler and more rapid than previous methods.
Fukuyama
R
R
Department of Pathology, Kyoto Prefectural University of Medicine, Japan.
Fushiki
S
S
Fujita
S
S
D000818:Animals; D001923:Brain Chemistry; D002417:Cattle; D002848:Chromatography, DEAE-Cellulose; D002852:Chromatography, Ion Exchange / Q000379:methods; D003598:Cytoskeletal Proteins / Q000302:isolation & purification; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005904:Glial Fibrillary Acidic Protein / Q000302:isolation & purification*; D015151:Immunoblotting; D008970:Molecular Weight; D010449:Peptide Mapping
D003598:Cytoskeletal Proteins; D005904:Glial Fibrillary Acidic Protein