Generation and Characterization of Monoclonal Antibodies That Specifically Recognize p65/L-Plastin Isoform but Not T-Plastin Isoform
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- TOYOOKA Kiminori
- International Medical Center of Japan
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- LIU Fenzhi
- Department of Immunology and Host Defenses, Ehime University School of Medicine
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- ISHII Motoshi
- Department of Immunology and Host Defenses, Ehime University School of Medicine
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- SAITO Shouichiro
- Department of Anatomy and Embryology, Ehime University School of Medicine
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- KIRIKAE Teruo
- International Medical Center of Japan
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- ASANO Yoshihiro
- Department of Immunology and Host Defenses, Ehime University School of Medicine
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- SHINOMIYA Hiroto
- Department of Immunology and Host Defenses, Ehime University School of Medicine
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Abstract
The 65-kDa protein (p65) was previously identified as a phosphorylated protein in activated macrophages, and has turned out to be a member of a plastin protein family characterized by a series of Ca2+-, calmodulin-, and β-actin-binding domains. In mice, two isoforms, p65/L-plastin and T-plastin, have so far been identified; p65/L-plastin is expressed in hemopoietic cells and cancer cells, and T-plastin in solid tissue cells. We generated monoclonal antibodies to p65/L-plastin, examined the isoform-specificity by using recombinant (r) T-plastin, and found that the antibodies were specific for rp65/L-plastin, whereas immune sera to rp65/L-plastin showed cross-reactions to rT-plastin. One of the antibodies, p65-7B5, was demonstrated to react to native p65/L-plastin by Western blot, flow cytometric, and immunohistochemical analysis. Furthermore, p65-7B5 has made it possible to detect p65/L-plastin-expressing cells in tissues where T-plastin is abundantly expressed. These reagents and procedures should provide specific tools to investigate the role of p65/L-plastin in leukocytes.
Journal
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- Bioscience, Biotechnology, and Biochemistry
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Bioscience, Biotechnology, and Biochemistry 70 (6), 1402-1407, 2006
Japan Society for Bioscience, Biotechnology, and Agrochemistry
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Details 詳細情報について
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- CRID
- 1390282681456276736
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- NII Article ID
- 10018530092
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- NII Book ID
- AA10824164
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- ISSN
- 13476947
- 09168451
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- NDL BIB ID
- 7958929
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- CiNii Articles
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- Abstract License Flag
- Disallowed