title: Application of Monoclonal Antibodies Against Mouse Dermokine.
abstract: Dermokine is one of the most highly expressed proteins in differentiating keratinocytes. Mouse dermokine has been reported to be encoded by 22 exons, and its expression leads to three transcripts, β, γ, and α, which are transcribed from two different transcriptional start sites. The α isoform represents the carboxyl-terminal domain of the β isoform, whereas the γ isoform lacks this domain. To reveal the distributions and expression levels of each isoform in mice, we generated rat monoclonal antibodies against dermokine-β/γ and dermokine-β/α. In immunofluorescence studies, the expression levels of dermokine in the cytosol of the cultured mouse keratinocytes were significantly elevated by high levels of extracellular calcium. In Western blot analyses, the expression levels of dermokine-β and dermokine-α were increased in the presence of high calcium. Finally, we developed a monoclonal antibody-based sensitive sandwich enzyme-linked immunosorbent assay (ELISA) and showed that the secreted dermokine-β into the culture medium from mouse keratinocytes was significantly increased in a manner dependent on the extracellular calcium concentration. These dermokine-specific antibodies have allowed us to gain new insights into the role of each dermokine isoform in cutaneous homeostasis.
authors.affiliation: 1 Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd. , Osaka, Japan .
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Higashi
authors.affiliation: 2 Department of Bioengineering, Graduate School of Engineering, Osaka City University , Osaka, Japan .
authors.forename: Cai-Xia
authors.initials: CX
authors.lastname: Wang
authors.affiliation: 3 Department of Biochemical Engineering, Graduate School of Science and Engineering, Yamagata University , Yamagata, Japan .
authors.forename: Chikako
authors.initials: C
authors.lastname: Yokoyama
authors.affiliation: 4 Laboratory of Toxicology, Faculty of Pharmacy, Osaka Ohtani University , Osaka, Japan .
authors.forename: Keita
authors.initials: K
authors.lastname: Yamada
authors.affiliation: 1 Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd. , Osaka, Japan .
authors.forename: Koichi
authors.initials: K
authors.lastname: Saito
authors.affiliation: 2 Department of Bioengineering, Graduate School of Engineering, Osaka City University , Osaka, Japan .
authors.forename: Taro
authors.initials: T
authors.lastname: Tachibana
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015153:Blotting, Western; D002118:Calcium / Q000494:pharmacology; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006367:HeLa Cells; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D015603:Keratinocytes / Q000166:cytology* / Q000276:immunology; D051379:Mice; D008810:Mice, Inbred C57BL; D020033:Protein Isoforms / Q000032:analysis / Q000096:biosynthesis* / Q000276:immunology; D011506:Proteins / Q000276:immunology*; D051381:Rats; D012867:Skin / Q000166:cytology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D036341:Intercellular Signaling Peptides and Proteins; D020033:Protein Isoforms; D011506:Proteins; C495569:dermokine protein, mouse; D002118:Calcium
keywords: cutaneous homeostasis; dermokine; isoform