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Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses
Podoplanin (PDPN) is known as a lymphatic endothelial cell marker. Monoclonal antibodies (mAbs) against human, mouse, rat, rabbit, dog, cat, bovine, pig, and horse PDPN have been established in our previous studies. However, mAbs against alpaca PDPN (aPDPN), required for immunohistochemical analysis...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6451175/ https://www.ncbi.nlm.nih.gov/pubmed/30997422 http://dx.doi.org/10.1016/j.bbrep.2019.100633 |
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author | Kato, Yukinari Furusawa, Yoshikazu Yamada, Shinji Itai, Shunsuke Takei, Junko Sano, Masato Kaneko, Mika K. |
author_facet | Kato, Yukinari Furusawa, Yoshikazu Yamada, Shinji Itai, Shunsuke Takei, Junko Sano, Masato Kaneko, Mika K. |
author_sort | Kato, Yukinari |
collection | PubMed |
description | Podoplanin (PDPN) is known as a lymphatic endothelial cell marker. Monoclonal antibodies (mAbs) against human, mouse, rat, rabbit, dog, cat, bovine, pig, and horse PDPN have been established in our previous studies. However, mAbs against alpaca PDPN (aPDPN), required for immunohistochemical analysis, remain to be developed. In the present study, we employed the Cell-Based Immunization and Screening (CBIS) method for producing anti-aPDPN mAbs. We immunized mice with aPDPN-overexpressing Chinese hamster ovary (CHO)-K1 cells (CHO/aPDPN), and hybridomas producing mAbs against aPDPN were screened using flow cytometry. One of the mAbs, PMab-225 (IgG(2b), kappa), specifically detected CHO/aPDPN cells via flow cytometry and recognized the aPDPN protein on Western blotting. Further, PMab-225 strongly stained lung type I alveolar cells, colon lymphatic endothelial cells, and kidney podocytes via immunohistochemistry. These findings demonstrate that PMab-225 antibody is useful to investigate the function of aPDPN via different techniques. |
format | Online Article Text |
id | pubmed-6451175 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-64511752019-04-17 Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses Kato, Yukinari Furusawa, Yoshikazu Yamada, Shinji Itai, Shunsuke Takei, Junko Sano, Masato Kaneko, Mika K. Biochem Biophys Rep Research Article Podoplanin (PDPN) is known as a lymphatic endothelial cell marker. Monoclonal antibodies (mAbs) against human, mouse, rat, rabbit, dog, cat, bovine, pig, and horse PDPN have been established in our previous studies. However, mAbs against alpaca PDPN (aPDPN), required for immunohistochemical analysis, remain to be developed. In the present study, we employed the Cell-Based Immunization and Screening (CBIS) method for producing anti-aPDPN mAbs. We immunized mice with aPDPN-overexpressing Chinese hamster ovary (CHO)-K1 cells (CHO/aPDPN), and hybridomas producing mAbs against aPDPN were screened using flow cytometry. One of the mAbs, PMab-225 (IgG(2b), kappa), specifically detected CHO/aPDPN cells via flow cytometry and recognized the aPDPN protein on Western blotting. Further, PMab-225 strongly stained lung type I alveolar cells, colon lymphatic endothelial cells, and kidney podocytes via immunohistochemistry. These findings demonstrate that PMab-225 antibody is useful to investigate the function of aPDPN via different techniques. Elsevier 2019-04-05 /pmc/articles/PMC6451175/ /pubmed/30997422 http://dx.doi.org/10.1016/j.bbrep.2019.100633 Text en © 2019 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Research Article Kato, Yukinari Furusawa, Yoshikazu Yamada, Shinji Itai, Shunsuke Takei, Junko Sano, Masato Kaneko, Mika K. Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses |
title | Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses |
title_full | Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses |
title_fullStr | Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses |
title_full_unstemmed | Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses |
title_short | Establishment of a monoclonal antibody PMab-225 against alpaca podoplanin for immunohistochemical analyses |
title_sort | establishment of a monoclonal antibody pmab-225 against alpaca podoplanin for immunohistochemical analyses |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6451175/ https://www.ncbi.nlm.nih.gov/pubmed/30997422 http://dx.doi.org/10.1016/j.bbrep.2019.100633 |
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