United Kingdom (UK)
Recombinant monoclonal antibody to Modified cytokeratin. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the human-human hybridoma COU-1.
UniProt Accession Number of Target Protein: P05787; P05783
Alternative Name(s) of Target: KRT8; CYK8; Keratin type II cytoskeletal 8; Cytokeratin-8; Type-II keratin Kb8; KRT18; CYK18; Keratin type I cytoskeletal 18; Cytokeratin-18; Cell proliferation-inducing gene 46 protein; COU1; B9165; ECACC 87040201
Immunogen: The original antibody was generated by fusing the human lymphoblastoid cell line WI-L2-729-HF2 with lymphocytes obtained from mesenteric lymph nodes from a colon cancer patient.
Specificity: This antibody binds modified forms of human cytokeratin 8 and cytokeratin 18. It is reported that this antibody recognizes an intracellularly located cytokeratin protein that is strongly expressed in adenocarcinomas.
Application Notes: This antibody reacted with a panel of colon cancer cell lines and melanoma cell lines in a immunocytochemical assay. This antibody strongly reacted with antigens expressed on autologous and allogeneic colorectal cancers in an immunohistochemical analysis of formalin-fixed paraffin-embedded tissues (PMID: 3699931). Electron microscopy of colonic adenocarcinoma cells, intact tumor and colonic epithelium by the immunogold technique demonstrated that the C-OU1 antibody reacted with a molecule associated with areas of disruption of the intermediate filaments in the cytoplasm of the tumor cells (PMID: 1607210). This antibody can also recognize modified cytokeratin from tumor extracts in a western blot (PMID: 1607210). This binding of this antibody to different recombinant heterotypic K8/K18 complexes was determined by ELISA. This antibody cannot bind K8(1-65)/intact K18 and intact K8(1-85)/intact K18 complexes in an ELISA (US20050048070). 131 Iodine labeled COU-1 antibody was also used in the detection and localization of tumors using immunoscintigraphy (PMID: 8306271; 8261404). The binding of this antibody to modified cytokeratin expressed on the surface of carcinoma cells causes their endocytosis, demonstrating that modified cytokeratin may serve as potential targets for delivery of immunoconjugates into cancerous cells (PMID: 9223323).
Antibody first published in:
Christensen et al. Human-human hybridomas for the study of anti-tumor immune response in patients with colorectal cancer. Int J Cancer. 1986 May 15;37(5):683-8. PMID:3699931
Note on publication:
Describes the generation of a human-human hybridoma with lymphocytes from patients with colorectal cancer.
Flow cytometry using anti-Modified cytokeratin antibody COU-1 (Ab03187). Paraformaldehyde fixed HeLa cells permeabilized with 0.5% Triton were stained with the anti-unknown specificity antibody (Ab00178-23.0; isotype control, black line) or the rabbit IgG version of COU-1 (Ab03187-23.0, blue line) at a dilution of 1:100 for 1h at RT. After washing, the bound antibody was detected using a goat anti-rabbit IgG AlexaFluor® 488 antibody at a dilution of 1:1000, and the cells were analyzed using a FACSCanto flow-cytometer.
Immunofluorescence staining of HeLa cells with anti-Modified cytokeratin antibody COU-1 (Ab03187). Immunofluorescence analysis of paraformaldehyde fixed HeLa cells, permeabilized with 0.15% Triton, on Shi-fix™ coverslips stained with the chimeric rabbit IgG version of COU-1 (Ab03187-23.0) (1:100 dilution) for 1h followed by Alexa Fluor® 488 secondary antibody (1:1000 dilution), showing cytoplasmic staining. The nuclear stain is DAPI (blue). Panels show, from left-right, top-bottom, Ab03187-23.0, DAPI, merged channels, and an isotype control. The isotype control was an unknown specificity antibody (Ab00178-23.0) followed by staining with Alexa Fluor® 488 secondary antibody.