The invention relates to an anti-SERINC5 antibody and application thereof. Specifically, the invention provides an anti-SERINC5 antibody and functional fragment, a nucleotide, a carrier, a monoclonalantibody, a kit and application thereof in detection of SERINC5 protein or polypeptide sequence shown as SEQIDNO:9. At the same time, the invention also provides a preparation method of the monoclonalantibody and a preparation method of a monoclonal antibody hybridoma cell line. The antibody and functional fragment prepared by the method provided by the invention can be effectively used for reaction with SERINC5 protein, like flow cytometry detection of SERINC5 molecules on cell surfaces and SERINC5 expressed in combination with cells, study of virus amplification mechanism, etc., thus providing an effective research tool for studying the antiviral mechanism of SERINC5, being conducive to accelerating the research progress in the field, and providing a new idea for further development ofnovel HIV protein Nef targeted AIDS drugs.