Critical Epitope of Anti-Rabbit Podoplanin Monoclonal Antibodies for Immunohistochemical Analysis

Ryusuke Honma, Yuki Fujii, Satoshi Ogasawara, Hiroharu Oki, Satoru Konnai, Yumiko Kagawa, Michiaki Takagi, Mika K. Kaneko, Yukinari Kato

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)65-72
Number of pages8
JournalMonoclonal antibodies in immunodiagnosis and immunotherapy
Volume35
Issue number2
DOIs
Publication statusPublished - 2016 Apr

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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