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Detection of Membrane‐Bound Complement Components by Crossed Immunoelectrophoresis of Target Membranes Solubilized with Non‐Ionic Detergent
Author(s) -
BHAKDI S.,
BJERRUM O. J.,
KNUFERMANN H.
Publication year - 1975
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.1365-3083.1975.tb03810.x
Subject(s) - immunoelectrophoresis , membrane , lysis , complement (music) , complement system , antibody , chemistry , membrane protein , solubilization , biochemistry , chromatography , biology , immunology , complementation , gene , phenotype
Sheep erythrocyte membranes lysed with human complement were solubilized in 1% Berol EMU‐043 (w/v) and subjected to crossed immunoelectrophoresis in the presence of this detergent. Using antibodies to human serum proteins, we found 5–6 protein precipitates whose presence was dependent on complement action. Among these, C3 and C4 components of complement were identified. Membrane‐bound C5 (C5b) did not precipitate in crossed immunoelectrophoresis, but its presence on target membranes was demonstrated by crossed‐line immunoelectrophoresis (in situ absorption) of specific anti‐CS antibodies. A reaction of partial identity between membrane‐bound C5 (CSb) and serum C5 was found. Our experimental approach allows immunoelectrophoretic analysis of complement components at the membrane level and should therefore prove useful for future investigations on the molecular structure of membrane‐bound complement.