Cell membrane coating with glutaraldehyde: application to a versatile solid-phase assay for thyroid membrane proteins and molecules interacting with thyroid membranes.
In defined conditions, glutaraldehyde was shown to tightly bind cell membranes to flexible microtiter plates without significant alteration of the antigenic and functional properties of membrane proteins. In the presence of 0.06% glutaraldehyde, human thyroid membranes were bound to plastic firmly enough to resist numerous washing and flicking steps; the coated membranes remained almost unaltered with regard to monoclonal antibody and thyrotropin binding as well as adenylate cyclase and peroxidase activities. Based on the use of thyroid membrane-coated microtiter plates, a versatile solid-phase assay was developed which allowed screening of anti-membrane monoclonal antibodies, detection of thyrotropin-displacing activity in hormone and antibody preparations, and monitoring of fractionation experiments of solubilized membrane antigens and thyrotropin receptor. It was concluded that the use of glutaraldehyde for coating cell membranes to flexible microtiter plates enabled the establishment of simple, rapid, and reliable assays for detection and quantitation of membrane proteins and molecules interacting with membranes.
D000262:Adenylyl Cyclases / Q000378:metabolism; D000447:Aldehydes; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000032:analysis; D002462:Cell Membrane / Q000378:metabolism; D005976:Glutaral; D006801:Humans; D007453:Iodide Peroxidase / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D008832:Microchemistry; D011956:Receptors, Cell Surface / Q000378:metabolism; D011989:Receptors, Thyrotropin; D012995:Solubility; D013961:Thyroid Gland / Q000378:metabolism*
D000447:Aldehydes; D000911:Antibodies, Monoclonal; D000941:Antigens; D008565:Membrane Proteins; D011956:Receptors, Cell Surface; D011989:Receptors, Thyrotropin; D007453:Iodide Peroxidase; D000262:Adenylyl Cyclases; D005976:Glutaral