Well-defined comb-on-comb copolymers of styrene, isoprene, and amethyl-styrene are prepared through cascade ''grafting-onto'' methods. The polymer main chain is prepared by nitroxide-mediated radical polymerization while the branches are prepared by anionic polymerization. The ''grafting-onto'' approach employs the coupling chemistry of macromolecular anions, such as polystyryllithium, polyisoprenyllithium, or poly(a-methylstyryl)lithium, toward either benzyl chloride or epoxy ring on precursor backbones. Thus a series of ABA-, ABB-, and ABC-type comb-on-comb copolymers are prepared and characterized by gel permeation chromatography equipped with a multi-angle laser light scattering detector and a viscometer. Unusual ''U-shaped'' dependences of radius of gyration, R g , on molecular weight are observed for comb-on-comb products, which are attributable to delayed elution of the densely grafted copolymers from GPC columns. The result also shows that the comb-on-comb copolymers exhibit morphologies from hard sphere to cylindrical rod, depending on the length ratio of the main chain to the branches. V V C 2008 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: [5518][5519][5520][5521][5522][5523][5524][5525][5526][5527] 2008