Monoclonal antibodies against rat NKp30 (rNKp30)

RNKp30 monoclonal antibodies were generated by immunizing BALB/c mice with rNKp30-Fc fusion protein. The rNKp30-Fc fusion protein is a soluble protein consisting of the extracellular domain of rNKp30 fused to the Fc domain of human IgG1. To produce rNKp30-Fc protein, we cloned a DNA construct fusing rNKp30 and human IgG1 Fc and stably transfected the DNA into 293F cells. RNKp30-Fc protein was secreted by 293F cells. Supernatants from these cells were harvested and the fusion protein was purified. Purified protein was used to inject into mice 3-4 times to generate an immune response. Serums and supernatants from hybridomas generated from the mice were screened for specificity to rNKp30 by ELISA and flow cytometry. Hybridomas positive for expression of rNKp30-specific antibodies were expanded. Supernatants from these hybridomas contain the anti-rNKp30 monoclonal antibodies.