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Objectives: ADP-Glc pyrophosphorylase(AGPase)is a key rate-limiting enzyme in maize starch biosynthesis,which exist in a heterotetrameric α2β2 structure in plant and is composed of Sh2 and Bt2 in maize endosperm.To understand AGPase subnuits protein expression in maize endosperm development,identification and preparation of maize AGPase subunit antibody.Methods:pGEX-6-t-GST-Sh2 and pGEX-6-t-GST-Bt2 expression vectors were constructed and transformed into Escherichia coli BL21(DE3).After induction with IPTG,protein waspurified by GST tag affinity purification method.Two healthy New Zealand white rabbits were immunized with Sh2 and Bt2 antigen mixed with freunds adjuvant respectively every two weeks until the antiserum were harvested.The specificity of antibodies were identified by western blot.Results: The results show that two polyclonal antibodies of Sh2 and Bt2 were specific torecognize the antigen and endogenous Sh2 and Bt2 protein from the maize endosperm.Protein expression model of Sh2 and Bt2 is similar in maize endosperm development.Both proteins have the highest expression level at 15 days after pollination(DAP).Conclusions: Polyclonal antibodies of Sh2 and Bt2 were successfully developed.The results from western blot show two polyclonal antibodies is specific to recognize endogenous Sh2and Bt2 protein in maize.As a good tool,both antibodies can be used in further functional research of AGPase.