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19. Detergent‐Containing Gels for Immunological Studies of Solubilized Erythrocyte Membrane Components
Author(s) -
BJERRUM OLE J.,
LUNDAHL PER
Publication year - 1973
Publication title -
scandinavian journal of immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.934
H-Index - 88
eISSN - 1365-3083
pISSN - 0300-9475
DOI - 10.1111/j.1365-3083.1973.tb03794.x
Subject(s) - solubilization , chemistry , precipitation , membrane , chromatography , immunoelectrophoresis , immunoprecipitation , ionic strength , membrane protein , staining , antibody , albumin , biochemistry , biology , immunology , aqueous solution , physics , genetics , meteorology , gene
Summary. Human erythrocyte membranes (ghosts) were dissolved to an extent of about 90% by repeated treatments at — 5° with the non‐ionic detergent Berol EMU‐043 in a concentration of 0.5% (w/v), and at pH 8.7. The solubilized membrane components were subjected to crossed immunoelectrophoresis with rabbit antibodies against ghost material. With 1% of Berol EMU‐043 in the gels, a recognizable pattern of 12 precipitation lines appeared. Only one of these was caused by a serum protein (albumin). The pattern of precipitates varied somewhat, probably due to some degradation of antigens by membrane‐bound proteolytic enzymes. Without detergent in the gels, a similar precipitation pattern was obtained, but some of the precipitation lines differed in shape and in staining intensity. In model experiments with human serum proteins, the incorporation of various non‐ionic detergents (Berol EMU‐043, Tween 20 SD, Triton X‐100, Lubrol WX, Nonidet P‐40) in a concentration of 2% (w/v) or less in the gels did not prevent immunoprecipitation of any protein.