Polyclonal and monoclonal antibodies specific for USP17, a proapoptotic deubiquitinating enzyme

USP17, a deubiquitinating enzyme (DUB), belongs to the ubiquitin-specific protease protein family. USP17 has highly conserved Cys, His, and Asp domains responsible for the deubiquitinating activity, and two hyaluronan binding motifs in its sequence. We reported that the presence of hyaluronan binding motifs in USP17 is responsible for the cell viability of cancerous cells. Although many if not all of the DUBs contain hyaluronan binding motifs in its sequence, the actual mechanism of hyaluronan binding motifs regulating signal transduction remains unclear. USP17 also regulates Ras activation and cell proliferation by inhibiting RCE1 activity. Thus USP17 may have a critical role in regulating tumor pathway through its deubiquitinating activity on various tumor-related proteins. This study reports the generation and characterization of polyclonal and six monoclonal antibodies against USP17 through immunoblotting, immunoprecipitation, and immunofluorescence microscopy analyses. These antibodies of USP17 will be useful as tools to investigate the mechanism of hyaluronan-mediated tumor suppression and also to screen interacting oncogenic substrates by immunoprecipitation assay.