Identification of a novel monoclonal antibody recognizing CD133.
Human CD133 (prominin-1), a cell surface glycoprotein, is used as a marker of hematopoietic and neural stem cells. Antibodies that recognize a glycosylation-dependent CD133 epitope have been extensively used for enrichment of tumor initiating cells in a variety of cancers. These currently available antibodies are restricted for use in only a subset of biological assays. We have generated a novel anti-human CD133 monoclonal antibody, using a recombinant protein consisting of highly immunogenic amino acid residues selected from the native CD133 protein as an immunogen. The antibody (identified as clone 7) specifically recognizes the CD133 protein in a variety of immunological applications including Western blot, immunofluorescence, flow cytometry and immunohistochemistry. Further, clone 7 specifically recognizes an unmodified CD133 extracellular domain, and not its glycosylated epitope. In conclusion, the specificity and usefulness in a wide range of applications suggest that clone 7 could be a valuable tool to identify CD133 positive cells as well as to target them for therapy.
Swaminathan
Suresh Kumar
SK
Department of Pharmaceutics, University of Minnesota, Minneapolis, MN 55455, USA.
Olin
Michael R
MR
Forster
Colleen L
CL
Cruz
Karen S Santa
KS
Panyam
Jayanth
J
Ohlfest
John R
JR
D000071916:AC133 Antigen; D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D015703:Antigens, CD / Q000276:immunology*; D015153:Blotting, Western; D018938:Caco-2 Cells; D000939:Epitopes / Q000276:immunology*; D005434:Flow Cytometry; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D014162:Transfection
D000071916:AC133 Antigen; D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D000939:Epitopes; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse