title: Critical Epitope of Anti-Rabbit Podoplanin Monoclonal Antibodies for Immunohistochemical Analysis.
abstract: Podoplanin (PDPN) is a type I transmembrane sialoglycoprotein, which is expressed in several normal cells, including lymphatic endothelial cells throughout the body, podocytes of the kidney, and lung type I alveolar cells of the lung. We have established many monoclonal antibodies (mAbs) against human PDPN, mouse PDPN, and rat PDPN. In addition, we recently produced an anti-rabbit PDPN mAb, PMab-32, which was established by immunizing mice with recombinant proteins of rabbit PDPN. Herein, we compared the reactivity of PMab-32 with that of newly established anti-rabbit PDPN mAbs, PMab-33 and PMab-21, against normal tissues in immunohistochemistry. PMab-32 reacted with podocytes, type I alveolar cells, and lymphatic endothelial cells, whereas PMab-33 detected only podocytes and type I alveolar cells but not lymphatic endothelial cells. PMab-21 was not useful in immunohistochemistry. We identified the epitope of PMab-32 and PMab-33 as Ser61-Ala68 of rabbit PDPN using western blot and flow cytometric analyses. In contrast, the epitope of PMab-21 was identified as Leu44-Glu48, which is corresponding to platelet aggregation-stimulating (PLAG) domain, indicating that Ser61-Ala68 of rabbit PDPN is a more appropriate epitope for immunohistochemistry compared with PLAG domain. PMab-32 could be useful for uncovering the function of rabbit PDPN.
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .|2 Department of Orthopaedic Surgery, Yamagata University Faculty of Medicine , Yamagata, Japan .
authors.forename: Ryusuke
authors.initials: R
authors.lastname: Honma
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Yuki
authors.initials: Y
authors.lastname: Fujii
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ogasawara
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .|2 Department of Orthopaedic Surgery, Yamagata University Faculty of Medicine , Yamagata, Japan .
authors.forename: Hiroharu
authors.initials: H
authors.lastname: Oki
authors.affiliation: 3 Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University , Hokkaido, Japan .
authors.forename: Satoru
authors.initials: S
authors.lastname: Konnai
authors.affiliation: 4 North Lab , Hokkaido, Japan .
authors.forename: Yumiko
authors.initials: Y
authors.lastname: Kagawa
authors.affiliation: 2 Department of Orthopaedic Surgery, Yamagata University Faculty of Medicine , Yamagata, Japan .
authors.forename: Michiaki
authors.initials: M
authors.lastname: Takagi
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Mika K
authors.initials: MK
authors.lastname: Kaneko
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Sendai, Japan .
authors.forename: Yukinari
authors.initials: Y
authors.lastname: Kato
mesh_terms: D056809:Alveolar Epithelial Cells / Q000276:immunology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D018604:Epitope Mapping; D000939:Epitopes / Q000276:immunology*; D057809:HEK293 Cells; D006801:Humans; D007150:Immunohistochemistry; D008562:Membrane Glycoproteins / Q000276:immunology*; D051379:Mice; D050199:Podocytes / Q000276:immunology; D011817:Rabbits; D051381:Rats; D013045:Species Specificity; D000098853:Podoplanin
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D008562:Membrane Glycoproteins; C117468:PDPN protein, human; D000098853:Podoplanin