Purification of the Thy-1 molecule from rat brain.
The present paper describes the characterization of the Thy-1 molecule from rat brain. The molecule was recognized by its antigens, which could be solubilized from brain membrane with deoxycholate. In the solubilized form the Thy-1 antigens were associated with a homogenous component with the following hydrodynamic properties: s20,w=2.2s,v=0.72ml/g and Stokes radius=3.0nm. The mol.wt. of the deoxycholateantigen complex was estimated to be 27000; these values are not significantly different from those obtained thymocyte Thy-1. Brain Thy-1 was further purified by affinity chromatography with lentil lectin coupled to Sepharose 4B, and more than 80% of the antigen was bound. The material eluted with methyl alpha-D-glucopyranoside was then filtered on a column of Sephadex G-200, and only one glycoprotein was found in the antigenically active fraction. On sodium dodecyl sulphate-polyacrylamide-gel electrophoresis the glycoprotein was very similar to the Thy-1 from thymocytes that binds to lentil lectin. Its apparent mol.wt. on 12.5% acrylamide gels was 24000, and it electrophoresed as a symmetrical band. Brain Thy-1 was antigenically indistinguishable from thymocyte Thy-1 when analysed with rabbit antisera raised against brain or thymocyte Thy-1.
D000818:Animals; D000906:Antibodies; D000941:Antigens; D001923:Brain Chemistry; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003840:Deoxycholic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D006023:Glycoproteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D037102:Lectins; D008566:Membranes; D011817:Rabbits; D051381:Rats; D012967:Sodium Dodecyl Sulfate; D012995:Solubility
D000906:Antibodies; D000941:Antigens; D000939:Epitopes; D006023:Glycoproteins; D037102:Lectins; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate