title: Development of Sensitive Monoclonal Antibody PMab-2 Against Rat Podoplanin.
abstract: Podoplanin (PDPN) is a platelet aggregation-inducing factor, which is known as an endogenous ligand of C-type lectin-like receptor-2 (CLEC-2). PDPN is also expressed in several normal tissues, such as lung type I alveolar cells and kidney podocytes. Although many monoclonal antibodies (MAbs) against human PDPN (hPDPN) or mouse PDPN (mPDPN) have been established, anti-rat PDPN (rPDPN) MAbs, especially against platelet aggregation-stimulating (PLAG) domain (29-54 amino acids) of rPDPN, have not been developed. Therefore, functional analysis of rPDPN in normal tissues has been limited. Here, we immunized mice with rPDPN peptides (38-51 amino acids) and developed a novel mouse anti-rPDPN MAb, PMab-2 (IgG1, kappa), which possesses high affinity compared with anti-hPDPN or mPDPN MAbs. The KD of PMab-2 was determined to be 5.9 × 10(-10) M. PMab-2 is useful, not only in flow cytometry and Western blot analysis against endogenous rPDPN, which is expressed in rat dermal fibroblast, but also in immunohistochemistry against normal tissues. PMab-2 showed extraordinarily high sensitivity in immunohistochemistry, indicating that PMab-2 is very advantageous for functional analysis of rPDPN.
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Aoba-ku, Sendai, Miyagi, Japan .|2 Department of Orthopaedic Surgery, Yamagata University Faculty of Medicine , Yamagata, Japan .
authors.forename: Hiroharu
authors.initials: H
authors.lastname: Oki
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Aoba-ku, Sendai, Miyagi, 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 , Aoba-ku, Sendai, Miyagi, Japan .
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ogasawara
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Aoba-ku, Sendai, Miyagi, Japan .
authors.forename: Yuki
authors.initials: Y
authors.lastname: Fujii
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Aoba-ku, Sendai, Miyagi, Japan .|2 Department of Orthopaedic Surgery, Yamagata University Faculty of Medicine , Yamagata, Japan .
authors.forename: Xing
authors.initials: X
authors.lastname: Liu
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 , Aoba-ku, Sendai, Miyagi, Japan .
authors.forename: Mika K
authors.initials: MK
authors.lastname: Kaneko
authors.affiliation: 1 Department of Regional Innovation, Tohoku University Graduate School of Medicine , Aoba-ku, Sendai, Miyagi, Japan .
authors.forename: Yukinari
authors.initials: Y
authors.lastname: Kato
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D000915:Antibody Affinity; D000918:Antibody Specificity; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D015870:Gene Expression; D006801:Humans; D007114:Immunization; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D007150:Immunohistochemistry / Q000379:methods*; D008562:Membrane Glycoproteins / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000008:administration & dosage / Q000138:chemical synthesis / Q000276:immunology; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D051381:Rats; D011994:Recombinant Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology; D000098853:Podoplanin
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D008562:Membrane Glycoproteins; D010455:Peptides; D011994:Recombinant Proteins; C092025:Pdpn protein, rat; D000098853:Podoplanin