Preparation of anti-NYD-SP8 rabbit polyclonal antibody and its application in the analysis of NYD-SP8 expression in nasopharyngeal carcinoma cell lines and clinical tissues

Tumori. 2011 Sep-Oct;97(5):655-9. doi: 10.1177/030089161109700519.

Abstract

Aims and background: NYD-SP8 is a recently identified protein, the biological characteristics of which are still unclear. The aim of this study was to prepare an anti-NYD-SP8 rabbit polyclonal antibody and investigate the expression of NYD-SP8 in human nasopharyngeal carcinoma cell lines and nasopharyngeal carcinoma tissues.

Methods: The anti-NYD-SP8 rabbit polyclonal antibody was prepared and ELISA was performed to assess the antibody titer. With this antibody, the NYD-SP8 expression in four nasopharyngeal carcinoma cell lines, CNE1, CNE2, 5-8F and 6-10B, was examined by Western blot and its expression in clinical tissues was also assessed by immunohistochemistry.

Results: The anti-NYD-SP8 rabbit polyclonal antibody with a high titer was successfully prepared. Western blot showed higher NYD-SP8 expression in CNE2 and 6-10B cells and lower expression in CNE1 and 5-8F cells. Immunohistochemistry demonstrated overexpression of NYD-SP8 in nasopharyngeal carcinoma tissue while the expression in normal nasopharyngeal tissue was negligible.

Conclusions: Our anti-NYD-SP8 rabbit polyclonal antibody can be used both for Western blot and immunohistochemistry, and can be a valuable tool to investigate the function and distribution of NYD-SP8. The different NYD-SP8 expression in various nasopharyngeal carcinoma cell lines indicated its complicated functions at different biological stages. The overexpression of NYD-SP8 in clinical nasopharyngeal carcinoma tissue indicated that it could play an important role in nasopharyngeal carcinoma carcinogenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies / analysis*
  • Biomarkers, Tumor / analysis*
  • Biomarkers, Tumor / immunology
  • Blotting, Western
  • Carcinoma / chemistry*
  • Carcinoma / pathology
  • Cell Line, Tumor
  • Enzyme-Linked Immunosorbent Assay
  • Gene Expression Regulation, Neoplastic
  • Humans
  • Immunohistochemistry
  • Membrane Proteins / analysis*
  • Membrane Proteins / immunology
  • Nasopharyngeal Carcinoma
  • Nasopharyngeal Neoplasms / chemistry*
  • Nasopharyngeal Neoplasms / pathology
  • Rabbits
  • Up-Regulation

Substances

  • Antibodies
  • Biomarkers, Tumor
  • Membrane Proteins
  • TEX101 protein, human