Aim To explore the preparation of the polyclonal antibody of a novel Homo sapiens synapse associated protein (FRG4). Methods FRG4 full-length sequence was obtained by PCR from human fetal liver library, by bioinformatics to detect the second structure of amino acids encoded by FRG4 and its epitope and motifs, according to the hydrophility, second structure, coupling and experiment difficulty etc, the peptide with 13 amino acid (PKLVKEEVFWRNY) was selected and synthesized by Solid-phase Peptide Synthesis (SSPS) method. After coupling,the peptide was immunized to rabbits through the basic and 6 fortified-immunities. 10 to 14 days later just after the fourth fortified-immunity,serum,separated from blood exsanguinated from carotid,was lyophilized and then stored at -20℃. Results Analyzed by ELISA and Western blot,the antibody dilution was 1∶16 000 and the blot was showed at the location of 40 kDa,the FRG4 protein was detected in cytoplasm of HepG2 cells by immunohistory. All these proved that the antibody has a good reaction and speciality. Experiments showed that the SSPS method was convenient to control the synthesis of peptides, spare time and labours,also obtain a good dilution. Conclusion A novel Homo sapiens synapse associated protein (FRG4) antibody was synthesized successfully,and worked as a foundation to the further study of FRG4 protein function playing in cardiovascular disease.