Immunohistochemical examination of novel rat monoclonal antibodies against mouse and human podoplanin

Chiaki Kaji, Yuta Tsujimoto, Mika Kato Kaneko, Yukinari Kato, Yoshihiko Sawa

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

This study aims to develop new monoclonal antibodies (mAbs) against mouse and human podoplanin. Rats were immunized with synthetic peptides, corresponding to amino acids 38-51 of mouse podoplanin or human podoplanin which is 100% homologous to the same site of monkey podoplanin; anti-mouse podoplanin mAb PMab-1 (IgG2a) and anti-human mAb NZ-1.2 (IgG2a) were established. In immunocytochemistry, the mouse melanoma B16-F10 and mouse podoplanin (mPDPN)-expressed CHO transfectant were stained by PMab-1; human lymphatic endothelial cells (LEC) and human podoplanin (hPDPN)-expressed squamous cell carcinoma HSC3 transfectant, were stained by NZ-1.2. Western-blot analysis detected an about 40-kDa protein in CHO-mPDPN and B16-F10 by PMab-1, and in HSC3-hPDPN and LEC by NZ-1.2. In frozen sections, PMab-1 reacted with mouse kidney, pulmonary alveoli, pulmonary pleura, and salivary gland myoepithelial cells while NZ-1.2 reacted to the human salivary gland myoepithelial cells. The immunostaining of paraffin-embedded sections also showed the reaction of PMab-1 or NZ-1.2 to the mouse or monkey kidney glomerulus, pulmonary alveoli, and lung lymphatic vessels. These results indicate that the two novel rat mAbs to the mouse and human/monkey podoplanin are useful for Western-blot and immunostaining of somatic tissues on paraffin-embedded sections as well as frozen sections.

Original languageEnglish
Pages (from-to)227-237
Number of pages11
JournalActa Histochemica et Cytochemica
Volume45
Issue number4
DOIs
Publication statusPublished - 2012
Externally publishedYes

Fingerprint

Endothelial cells
Paraffin
Rats
Monoclonal Antibodies
Tissue
Pulmonary Alveoli
Amino Acids
Peptides
Haplorhini
Frozen Sections
Salivary Glands
Proteins
Endothelial Cells
Western Blotting
Kidney Glomerulus
Lung
Experimental Melanomas
Lymphatic Vessels
Pleura
Squamous Cell Carcinoma

Keywords

  • Monoclonal antibody
  • NZ-1.2
  • PMab-1
  • Podoplanin

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Physiology
  • Pathology and Forensic Medicine
  • Histology

Cite this

Immunohistochemical examination of novel rat monoclonal antibodies against mouse and human podoplanin. / Kaji, Chiaki; Tsujimoto, Yuta; Kaneko, Mika Kato; Kato, Yukinari; Sawa, Yoshihiko.

In: Acta Histochemica et Cytochemica, Vol. 45, No. 4, 2012, p. 227-237.

Research output: Contribution to journalArticle

Kaji, Chiaki ; Tsujimoto, Yuta ; Kaneko, Mika Kato ; Kato, Yukinari ; Sawa, Yoshihiko. / Immunohistochemical examination of novel rat monoclonal antibodies against mouse and human podoplanin. In: Acta Histochemica et Cytochemica. 2012 ; Vol. 45, No. 4. pp. 227-237.
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