To circumvent this problem, we developed a simple two-step procedure consisting of first the deposition of a quasi-hexagonal pattern of homogeneously distributed gold nanoparticles (AuNPs) by block-copolymer micellar lithography on ITO. Subsequently, the AuNPs act as seeds for further gold growth by electrodeposition on the pre-patterned ITO substrate. In this way, we reproducibly achieved substrates (>50) with an average gold coverage of (40 ± 5) % and many hot spots due to a small average inter-particle distance of (15 ± 5) nm. These substrates exhibit a strong and homogeneous Raman signal, as determined using 2D maps obtained with the standard Raman tag molecule 4-mercaptobenzoic acid. Moreover, the electrochemical performance of the developed conductive SERS substrates was demonstrated using a Michael addition reaction monitored by Raman scattering. This reaction occurs between the hydroquinone/benzoquinone redox pair and a biologically relevant analyte, but only takes place in one of the redox states of the hydroquinone/benzoquinone system.