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This paper describes a new sensitive microELISA based on enzyme/anti-enzyme complexes following an unlabelled antibody bridging step. beta-Galactosidase/anti-beta-galactosidase complexes were made using a monoclonal antibody raised against bacterial (E. coli) beta-galactosidase and enzyme activity was quantified with a fluorogenic substrate. Because of its high sensitivity the assay is particularly suitable for the detection of limited amounts of antigen. One application illustrated is the analysis of Class I and Class II histocompatibility antigens on peripheral blood lymphocytes using 5000 cells/well in 60-well Terasaki or 96-well microtitre plates.

Original publication

DOI

10.1016/0022-1759(87)90100-1

Type

Journal article

Journal

J Immunol Methods

Publication Date

26/02/1987

Volume

97

Pages

19 - 27

Keywords

Animals, Antibodies, Monoclonal, Antigen-Antibody Complex, Escherichia coli, Galactosidases, Immunoassay, Mice, Mice, Inbred BALB C, Microchemistry, beta-Galactosidase