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

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.