Biomaterial Database

Monoclonal antibodies to soluble, human milk galactosyltransferase (lactose synthase A protein). 1986

E G Berger, and E Aegerter, and T Mandel, and H P Hauri

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.

UI	MeSH Term	Description	Entries
D007700	Kinetics	The rate dynamics in chemical or physical systems.
D007788	Lactose Synthase	An enzyme complex that catalyzes the transfer of GALACTOSE from UDP GALACTOSE to GLUCOSE, forming LACTOSE. The enzyme complex is composed of a B subunit, ALPHA-LACTALBUMIN, which changes the substrate specificity of the A subunit, N-ACETYLLACTOSAMINE SYNTHASE, from N-ACETYLGLUCOSAMINE to glucose making lactose synthesis the preferred reaction.	Lactose Synthetase,N-Acetylgucosamide 1-4 beta Galactosyl Transferase,N Acetylgucosamide 1 4 beta Galactosyl Transferase,Synthase, Lactose,Synthetase, Lactose
D008895	Milk, Human	Milk that is produced by HUMAN MAMMARY GLANDS.	Breast Milk,Human Milk,Milk, Breast
D009237	N-Acetyllactosamine Synthase	The A protein of the lactose synthase complex. In the presence of the B protein (LACTALBUMIN) specificity is changed from N-acetylglucosamine to glucose. EC 2.4.1.90.	N-Acetyllactosamine Synthetase,UDP Galactose Acetylglucosamine Galactosyltransferase,N-Acetylglucosamine beta-D-Galactosyltransferase,UDP Galactose-N-Acetylglucosamine Galactosyltransferase,beta-1,4-Galactosyltransferase,Galactosyltransferase, UDP Galactose-N-Acetylglucosamine,N Acetylglucosamine beta D Galactosyltransferase,N Acetyllactosamine Synthase,N Acetyllactosamine Synthetase,Synthase, N-Acetyllactosamine,Synthetase, N-Acetyllactosamine,UDP Galactose N Acetylglucosamine Galactosyltransferase,beta 1,4 Galactosyltransferase,beta-D-Galactosyltransferase, N-Acetylglucosamine
D011247	Pregnancy	The status during which female mammals carry their developing young (EMBRYOS or FETUSES) in utero before birth, beginning from FERTILIZATION to BIRTH.	Gestation,Pregnancies
D002999	Clone Cells	A group of genetically identical cells all descended from a single common ancestral cell by mitosis in eukaryotes or by binary fission in prokaryotes. Clone cells also include populations of recombinant DNA molecules all carrying the same inserted sequence. (From King & Stansfield, Dictionary of Genetics, 4th ed)	Clones,Cell, Clone,Cells, Clone,Clone,Clone Cell
D004797	Enzyme-Linked Immunosorbent Assay	An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed.	ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D005260	Female		Females
D005455	Fluorescent Antibody Technique	Test for tissue antigen using either a direct method, by conjugation of antibody with fluorescent dye (FLUORESCENT ANTIBODY TECHNIQUE, DIRECT) or an indirect method, by formation of antigen-antibody complex which is then labeled with fluorescein-conjugated anti-immunoglobulin antibody (FLUORESCENT ANTIBODY TECHNIQUE, INDIRECT). The tissue is then examined by fluorescence microscopy.	Antinuclear Antibody Test, Fluorescent,Coon's Technique,Fluorescent Antinuclear Antibody Test,Fluorescent Protein Tracing,Immunofluorescence Technique,Coon's Technic,Fluorescent Antibody Technic,Immunofluorescence,Immunofluorescence Technic,Antibody Technic, Fluorescent,Antibody Technics, Fluorescent,Antibody Technique, Fluorescent,Antibody Techniques, Fluorescent,Coon Technic,Coon Technique,Coons Technic,Coons Technique,Fluorescent Antibody Technics,Fluorescent Antibody Techniques,Fluorescent Protein Tracings,Immunofluorescence Technics,Immunofluorescence Techniques,Protein Tracing, Fluorescent,Protein Tracings, Fluorescent,Technic, Coon's,Technic, Fluorescent Antibody,Technic, Immunofluorescence,Technics, Fluorescent Antibody,Technics, Immunofluorescence,Technique, Coon's,Technique, Fluorescent Antibody,Technique, Immunofluorescence,Techniques, Fluorescent Antibody,Techniques, Immunofluorescence,Tracing, Fluorescent Protein,Tracings, Fluorescent Protein
D006367	HeLa Cells	The first continuously cultured human malignant CELL LINE, derived from the cervical carcinoma of Henrietta Lacks. These cells are used for, among other things, VIRUS CULTIVATION and PRECLINICAL DRUG EVALUATION assays.	Cell, HeLa,Cells, HeLa,HeLa Cell