Herein, we report a novel method for H2O2 detection based on a single plasmonic nanoprobe via cytochrome c (Cyt c)-mediated plasmon resonance energy transfer (PRET). Dynamic spectral changes were observed in the fingerprint quenching dip of a single plasmonic nanoprobe in response to changes in the redox state of Cyt c, induced by H2O2. Based on the changes in the spectral profile of the single plasmonic nanoprobe, H2O2 was successfully detected in a wide concentration range from 100 mM to 10 nM, including physiologically relevant micromolar and nanomolar concentrations.