Nitrogen addition may alter the decomposition rate for different organic-matter pools in contrasting ways. Using a paired-plot design, we sought to determine the effects of long-term elevated N on the stability of five organic-matter pools: organic horizons (Oe+a), whole mineral soil (WS), mineral soil fractions including the light fraction (LF), heavy fraction (HF), and a physically recombined fraction (RF). These substrates were incubated for 300 days, and respiration, mineralized N, and active microbial biomass were measured. Samples with elevated N gave 15% lower cumulative respiration for all five substrates. Over the 300-day incubation, the Oe+a gave twice the cumulative respiration (g C kg−1 initial C) as the LF, which gave slightly higher respiration than the HF. Respiration was 35% higher for the WS than for the RF. Mineralized N was similar between N treatments and between the LF and HF. Net N mineralized by the LF over the course of the 300-day incubation decreased with higher C:N ratio, due presumably to N immobilization to meet metabolic demands. The pattern was opposite for HF, however, which could be explained by a release of N in excess of metabolic demands due to recalcitrance of the HF organic matter. Mineralized N increased with respiration for the HF but showed no pattern, or perhaps even decreased, for the LF. WS and RF showed decreasing active microbial biomass near the end of the incubation, which corresponded with decreasing respiration and increasing nitrate. Our results show that long-term elevated N stabilized organic matter in whole soil and soil fractions.