Ag/Au bimetallic nanoparticles (BNPs) with a., size less than 2 nm were prepared by physical mixture of colloidal dispersions of Ag and Au nanoparticles (NPs). This provides an example of fabrication of BNPs with self-organization by the reaction between metal NPs. Although Ag/Au BNPs having different structures and compositions are one of the most widely studied bimetallic systems in the literature due to their wide range of uses such as in catalysis, electronics, plasmonics, optical sensing, and surface-enhanced Raman scattering, we first prepared such BNPs by physical mixture and characterized them by UV-vis spectroscopy, SERS, XPS, TEM, and EDS in HR-STEM. The present fabrication method has the advantage of avoiding the unfavorable formation of AgCl precipitates in the reaction process which are always produced when Ag+ ions are used as a starting material in combination with a HAuCl4 precursor. These Ag/Au BNPs showed high catalytic activities for aerobic glucose oxidation, and the highest activity of 11 510 mol of glucose.h(-1).mol of metal(-1) was observed for the BNPs with a Ag/Au atomic ratio of 1/4; the activity value is about 2 times higher than that of Au NPs with nearly the same particle size. XPS and DFT calculation results show that the negatively charged Au atoms due to the electron charge transfer effects from neighboring Ag atoms and poly(N-vinyl-2-pyrrolidone) act as catalytically active sites and play an important role in the aerobic glucose oxidation.