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Monoclonal antibodies to soluble, human milk galactosyltransferase (lactose synthase A protein)

Berger, E. G. and Aegerter, E. and Mandel, T. and Hauri, H. P.. (1986) Monoclonal antibodies to soluble, human milk galactosyltransferase (lactose synthase A protein). Carbohydrate research, Vol. 149, H. 1. pp. 23-33.

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Official URL: http://edoc.unibas.ch/dok/A5257830

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Abstract

Monoclonal antibodies have been produced against soluble human milk galactosyltransferase of a blood group O donor. After initial screening by radioimmunoassay, fourteen hybridomas were further characterized by enzyme-linked immunosorbent assay, immunoblotting of purified enzyme following sodium dodecyl sulfate-polyacrylamide gel electrophoresis, enzyme activity modification, and enzyme localization in HeLa cells by immunofluorescence. Of these fourteen clones, seven had titers between 1500 and 7800 as estimated by ELISA. In general, the titer correlated with staining intensity on immunoblots and in immunofluorescence. In the presence of monoclonal antibody, enzyme activity was usually slightly enhanced or stabilized. Subcloning yielded four monoclonal antibody preparations designated as GT2/24/108, GT2/36/118, GT2/61/14, and GT2/77/22, which belong to Ig class G2b, G3, M, and G1, respectively. They all recognized the enzyme in purified form or in defatted milk as a single, broad band on electrophoresis-immunoblotting and produced a concise juxtanuclear fluorescence typical for the Golgi apparatus in HeLa cells.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Former Organization Units Biozentrum > Pharmacology/Neurobiology (Hauri)
UniBasel Contributors:Hauri, Hans-Peter
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Elsevier Scientific Publishing
ISSN:0008-6215
Note:Publication type according to Uni Basel Research Database: Journal article
Last Modified:22 Mar 2012 14:22
Deposited On:22 Mar 2012 13:29

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