OBJECTIVE Prepare the rabbit antiserum against gecko japonicus Hoxc10 and to identify its properties. METHODS Prokaryotic expression vector of g-Hoxc10 were constructed and then transform into E.coli (BL21). To make GST-g-Hoxc10 fusion protein in E.coli (BL21) under the optimized induction of Isopropyl β-D-1-thiogalactopyranoside(IPTG). The recombination proteins were purified using affinity chromatography. The purified fusion protein was inoculated into adult rabbits to develop antiserum. Western blot and immunohistochemistry staining were then performed to evaluate the feature of the prepared antiserum. RESULTS Prokaryotic expression vectors of g-Hoxc10 were successfully constructed. The soluble recombinant protein was highly expressed in E.coli BL21 and inoculated into adult rabbits to obtain high titer antiserum. Western blot and immunohistochemistry staining were then performed to evaluate the specificity of the prepared antiserum. CONCLUSIONS We successfully amplified and expressed the g-Hoxc10 in E.coli BL21. The purified fusion protein was inoculated into adult rabbits to develop antiserum. The obtained antiserum of g-Hoxc10 showed a high titer against Hoxc10 proteins. The protein and antiserum prepared in this study can be used for further research of the function investigation of Hoxc10.