Generation of Rat Monoclonal Antibody for Human Nucleolin

Nucleolin (NCL) is a multifunctional phosphoprotein that is mainly localized in the nucleolus, but it is also found in the nucleoplasm, cytoplasm, and cell membrane. The principal functions of NCL involve DNA and RNA metabolism, gene transcription and translation, ribosome biogenesis, and mRNA stability. It was also reported that the localization of human NCL (hNCL) is related to tumor malignancy. Therefore, analyzing the cellular dynamics of NCL could be useful. In this article, we describe rat monoclonal antibody (mAb) 6F9A6 that was generated against a hNCL peptide. This mAb recognizes endogenous human, monkey, dog, and mouse NCL and was shown to be useful in immunofluorescence staining, immunoprecipitation, and immunoblotting experiments in several cancer cell lines. We anticipate that the mAb 6F9A6 will be useful for functional analyses of hNCL in cancer cells.