A flow cytometric screening test for detergent-resistant surface antigens in monocytes.
Rafts resemble cholesterol- and glycosphingolipid-enriched, liquid-ordered plasma membrane microdomains, showing resistance to nonionic detergents, and are involved in various cellular processes. In the present study, we have tested surface antigens on resting and lipopolysaccharide (LPS)-stimulated human peripheral blood monocytes for their detergent resistance (i.e. raft-association), by flow cytometry. Constitutive (CD14, CD32, CD55), or LPS-induced (CD81) raft-association, and detergent solubility (i.e. exclusion of rafts) (CD71) of monocyte antigens in the presence of 0.01% Triton X-100 are clearly demonstrated. Flow cytometric detergent insolubility is a powerful tool for rapid screening the raft-association of monocyte antigens in a whole-blood assay.
Wolf
Zsuzsanna
Z
Institute for Clinical Chemistry and Laboratory Medicine, University of Regensburg, Regensburg, Germany.
Orsó
Evelyn
E
Werner
Tobias
T
Boettcher
Alfred
A
Schmitz
Gerd
G
D015703:Antigens, CD / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis* / Q000378:metabolism; D018958:CD55 Antigens / Q000032:analysis; D002462:Cell Membrane / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D003902:Detergents / Q000737:chemistry*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007700:Kinetics; D018950:Lipopolysaccharide Receptors / Q000032:analysis; D008070:Lipopolysaccharides / Q000494:pharmacology; D021962:Membrane Microdomains / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D009000:Monocytes / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D017830:Octoxynol / Q000737:chemistry; D017452:Receptors, IgG / Q000032:analysis; D011990:Receptors, Transferrin / Q000032:analysis; D012995:Solubility; D060225:Tetraspanin 28
D015703:Antigens, CD; D000954:Antigens, Surface; D018958:CD55 Antigens; C068126:CD71 antigen; C554798:CD81 protein, human; D003902:Detergents; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D017452:Receptors, IgG; D011990:Receptors, Transferrin; D060225:Tetraspanin 28; D017830:Octoxynol