We use the antenna subtraction method to isolate the mixed real-virtual infrared singularities present in gluonic scattering amplitudes at next-to-next-to-leading order. In a previous paper, we derived the subtraction term that rendered the double real radiation tree-level process finite in the single and double unresolved regions of phase space. Here, we show how to construct the real-virtual subtraction term using antenna functions with both initial-and final-state partons which removes the explicit infrared poles present in the one-loop amplitude, as well as the implicit singularities that occur in the soft and collinear limits. As an explicit example, we write down the subtraction term that describes the single unresolved contributions from the five-gluon one-loop process. The infrared poles are explicitly and locally cancelled in all regions of phase space prior to integration, leaving a finite remainder that can be safely evaluated numerically in four-dimensions. We show numerically that the subtraction term correctly approximates the matrix elements in the various single unresolved configurations.