Electrochemiluminescence (ECL) detection technique using bipyridine-ruthenium(ii) complexes as probes is a highly sensitive and widely used method for the detection of various biological and bioactive molecules. In this work, the spectral, electrochemical and ECL properties of a chemically modified bipyridine-ruthenium(ii) complex, [Ru(bpy)2(dabpy)] 2+ (bpy: 2,2′-bipyridine; dabpy: 4-(3,4-diaminophenoxy)-2, 2′-bipyridine), were investigated and compared with those of its nitric oxide (NO)-reaction derivative [Ru(bpy)2(T-bpy)]2+ (T-bpy: 4-triazolephenoxy-2,2′-bipyridine) and [Ru(bpy)3] 2+. It was found that the ECL intensity of [Ru(bpy) 2(dabpy)]2+ could be selectively and sensitively enhanced by NO due to the formation of [Ru(bpy)2(T-bpy)]2+ in the presence of tri-n-propylamine. By using [Ru(bpy)2(dabpy)] 2+ as a probe, a sensitive and selective ECL method with a wide linear range (0.55 to 220.0 μM) and a low detection limit (0.28 μM) was established for the detection of NO in aqueous solutions and living cells. The results demonstrated the utility and advantages of the new ECL probe for the detection of NO in complicated biological samples.