Monoclonal antibodies recognizing different epitopes of cytokeratin No.18.
A comparative study of six mouse monoclonal antibodies against human 45 kDa keratin polypeptide (keratin No. 18) was undertaken using three experimental approaches: immunohistochemistry on normal human tissues, examination of interspecies cross-reactivity and identification of the target polypeptides in 1-D and 2-D immunoblots. The data suggest that at least five different antigenic sites of keratin 18 are recognized by this panel of reagents. The C-04 epitope is keratin 18-specific and widely conserved among mammalian species, while the antibodies DA7 and DC10 also react specifically with the 45 kDa keratin but stain simple epithelia of human origin only. Two antibodies, C-11 and C-66, decorate simple as well as stratified epithelia in human and in all seven animal species tested, but their respective target epitopes are shared by different groups of keratin polypeptides, which indicates their non-identity. In contrast to keratin specificity of the five above mentioned antibodies, the C-08 antibody cross-reacts with a 70 kDa nuclear lamina protein found in human and bovine tissues. The results of the present study provide the necessary basis for future applications of these antibodies in both routine immunodiagnostic work and as probes to study the biology of epithelial cells in general and the significance of keratin intermediate filaments in particular.
Vojtĕsek
B
B
Department of Immunology and Molecular Biology, Research Institute of Clinical and Experimental Oncology, Brno.
Stasková
Z
Z
Nenutil
R
R
Lauerová
L
L
Kovarík
J
J
Rejthar
A
A
Bártková
J
J
Bártek
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002417:Cattle; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000276:immunology; D004848:Epithelium / Q000276:immunology; D000939:Epitopes / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D015151:Immunoblotting; D007150:Immunohistochemistry; D007633:Keratins / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D013045:Species Specificity; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007633:Keratins