title: 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: OBJECTIVE
  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.
authors.affiliation: 1State Key Laboratory of Oncology in South China, Guangzhou, Guangdong, China.
authors.forename: Ling-Lin
authors.initials: LL
authors.lastname: Yang
authors.forename: Jing-Bo
authors.initials: JB
authors.lastname: Wu
authors.forename: Zhi-Gang
authors.initials: ZG
authors.lastname: Liu
authors.forename: Li
authors.initials: L
authors.lastname: U
authors.forename: Mu-Sheng
authors.initials: MS
authors.lastname: Zeng
authors.forename: Shan-Shan
authors.initials: SS
authors.lastname: Yan
authors.forename: Hsiao-Chang
authors.initials: HC
authors.lastname: Chan
authors.forename: Yun-Fei
authors.initials: YF
authors.lastname: Xia
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis*; D014408:Biomarkers, Tumor / Q000032:analysis* / Q000276:immunology; D015153:Blotting, Western; D002277:Carcinoma / Q000737:chemistry* / Q000473:pathology; D045744:Cell Line, Tumor; D004797:Enzyme-Linked Immunosorbent Assay; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000032:analysis* / Q000276:immunology; D000077274:Nasopharyngeal Carcinoma; D009303:Nasopharyngeal Neoplasms / Q000737:chemistry* / Q000473:pathology; D011817:Rabbits; D015854:Up-Regulation
chemical_list: D000906:Antibodies; D014408:Biomarkers, Tumor; D008565:Membrane Proteins; C539319:TEX101 protein, human