Production and characterization of monoclonal antibodies that recognize bovine Kit receptor.
Kit receptor is a transmembrane tyrosine kinase that is the receptor for stem cell factor (SCF). The extracellular domain of bovine Kit receptor (boKit) was produced by a baculovirus expression system. Six monoclonal antibody (MAb) clones designated as bK-1 to bK-6 were obtained upon immunization of mice with the recombinant protein. Immunoprecipitation and flow cytometric analysis indicated that all of the MAbs specifically bound to boKit expressed in COS-7 cells transfected with boKit cDNA. Four of the six MAbs neutralized the biological activity of recombinant bovine SCF, whereas the other two did not. The boKit-positive and boKit-negative cell fractions were sorted from cryopreserved bovine bone marrow cells by the use of MAb bK-1. Colony formation assays indicated that the cells which were able to grow in response to bovine SCF were enriched in the boKit-positive fraction. These MAbs would be valuable in studying possible boKit-positive cell species such as bovine hematopoietic cells, and in defining the biological role of Kit receptor in cattle.
Hikono
H
H
National Institute of Animal Health, Tsukuba, Ibaraki, Japan.
Ohta
M
M
Kubota
T
T
Zhou
J H
JH
Inumaru
S
S
Sakurai
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000096:biosynthesis*; D000918:Antibody Specificity; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology; D019556:COS Cells; D002417:Cattle / Q000276:immunology*; D017931:DNA Primers / Q000737:chemistry; D018604:Epitope Mapping / Q000662:veterinary; D000939:Epitopes / Q000276:immunology; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary; D051379:Mice; D008807:Mice, Inbred BALB C; D011233:Precipitin Tests / Q000662:veterinary; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics / Q000276:immunology*; D019089:Stem Cell Factor / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D000939:Epitopes; D019089:Stem Cell Factor; D019009:Proto-Oncogene Proteins c-kit