This work aims to experimentally clarify how the single chain conformation evolves as a function of grafting density for model comb-like chains in dilute solution in the whole density regime. Via a combination of rational design, precise synthesis and accurate characterization, we obtained four sets of PPANb-g-PS30-σ comb-like samples with well-defined architectures and accurately extracted their molecular parameters by triple detection size exclusion chromatography (TD-SEC). With these samples in hand, we quantified how the excluded volume interaction and chain conformation evolve with the grafting density (σ) in the whole density regime. Three main findings are reported: (i) the graft-graft excluded volume interaction is not ignorable even in the low σ-regime; (ii) contrary to theoretical predictions, both the excluded volume interaction and the chain conformation are found to be Nb-dependent; (iii) both Rg ∼ σ1/3 and [η] ∼ σ0 are experimentally confirmed for comb-like chains from different σ and Nb, signifying a unique 3D mass-size growth pattern and a quasi-3D fractal feature. The obtained results help clarify some long-existed controversial issues in the field.