Astragalus polysaccharide (APS) was obtained by hot water extraction, alcohol precipitation, gel-permeation chromatography and ultrafiltration. Fluorescence material 2-aminoacridone (2-AMAC) labeled APS bind to macrophage in a time- dependent manner and the binding can be remarkably inhibited by APS. Furthermore, the effect of APS on RAW264.7 macrophage demonstrated APS increase the level of cytokines including TNF-α, GM-CSF and the production of NO. NF-κB protein levels are increased in response to APS. Blocking NF-κB with specific inhibitor resulted in decreased levels of NO and TNF-α. The results suggested that APS possess potent immunomodulatory activity by stimulating macrophage and could be used as an immunotherapeutic adjuvant.