Covalent attachment of enzyme as a membrane-label for viable eucaryotic cells.
An enzyme, beta-D-galactosidase, was covalently coupled to mammalian cells by means of a bifunctional reagent. The coupling procedure did not cause appreciable loss of cell viability (less than 6%) as measured by plating efficiently and membrane integrity. After 24 h in culture, the cells exhibited an average of 2.6 x 10(4) molecules of beta-D-galactosidase per cell. Histological evidence indicated that the enzyme was localized on the cell surface and distributed uniformly among the cell population. Considerations for choosing enzyme-label include sensitivity of assay by enzymatic, immunologic and histochemical methods, and the possibility of isolating labeled membrane components by enzyme-specific affinity chromatography.
D000818:Animals; D001345:Autoradiography; D002462:Cell Membrane / Q000032:analysis* / Q000378:metabolism; D002470:Cell Survival; D002478:Cells, Cultured; D003584:Cytological Techniques; D005696:Galactosidases; D051379:Mice; D001616:beta-Galactosidase / Q000378:metabolism
D005696:Galactosidases; D001616:beta-Galactosidase