Abstract:
Protein tyrosine kinases (PTKs)-mediated signaling transduction pathways play pivotal roles in the regulation of key functions during embryogenesis, development, metabolism and immunity.PTKs enhance the cell proliferation, mobility and viability by activating multiple downstream signaling transduction pathways, whereas abnormal PTKs catalytic activity often leads to oncogenesis.NOK/STYK1（novel oncogene with kinase domain, also known as serine/threonine and tyrosine receptor protein kinase）is a new member of PTKs that has been identified as another oncogene.However, the mechanism of NOK/STYK1 participates in the regulation of signaling transduction and oncogenesis remains elusive.Thus, to generate a specific monoclonal antibody to illustrate the functions of NOK/STYK1 is really in need.In this research, we designed and purified some short peptides of NOK/STYK1 as antigens to immunize Balb/C mice to obtain the B cells of spleen.Subsequently, we fused the B cells to murine cells SP2/0 and then screened and identified the positive hybridoma cells line for production of the monoclonal antibody against NOK/STYK1 by ELISA.Fortunately, we found that a hybridoma cell line 7G9 has a high production of monoclonal antibody, which was confirmed for the specificity and efficiency by both western blotting and ELISA.This monoclonal antibody was verified to be powerful for detecting the expression of NOK/STYK1 in various cancer cell lines and tissues by both immunefluorescence and immunohistochemical staining respectively.
