Azo dyes are a class of photoactive dyes that constitute a major focus of chemical research due to their applications in numerous industrial functions. This work explores the impact of protonation on the photophysics of four naphthalene-based azo dyes. The pKa value of the dyes increases proportionally with decreasing Hammett parameter of p-phenyl substituents from 8.1 (R = -H, σ = 0) to 10.6 (R = -NMe2, σ = -0.83) in acetonitrile. Protonation of the dyes shuts down the steady-state photoisomerization observed in the unprotonated moieties. Fluorescence measurements reveal a lower quantum yield with more electron-donating p-phenyl substituents, with overall lower fluorescence quantum yields than the unprotonated dyes. Transient absorption spectroscopy reveals four excited-state lifetimes (<1 ps, ∼3 ps, ∼13 ps, and ∼200 ps) exhibiting faster excited-state dynamics than observed in the unprotonated forms (for 1-3: 0.7-1.5 ps, ∼3-4 ps, 20-40 ps, 20-300 min; for 4: 0.7 ps, 4.8 ps, 17.8 ps, 40 ps, 8 min). Time-dependent density functional theory (TDDFT) elucidates the reason for the loss of isomerization in the protonated dyes, revealing a significant change in the lowest excited state potential energy nature and landscape upon protonation. Protonation impedes relaxation along the typical rotational and inversion isomerization axes, locking the dyes into a trans-configuration that rapidly decays back to the ground state.