This study focused on the experimental assessment of the effect of the spacing between longitudinal stiffeners welded to I-shaped beams under the action of lateral-torsional buckling. In this procedure, 192 aluminum beams on a 1:9 scale were tested under simple-support conditions with a laterally unbraced length ranging from 0.55 m through 1.95 m. Moreover, the stiffeners’ spacing was also ranged from 3 to 9 times the depth of section. The structural behavior of the beams is discussed in terms of their flexural capacity, spacing between longitudinal stiffeners, lateral displacement of compression flange and failure angle twist. Results show that the spacing of longitudinal stiffeners influences the flexural capacity of I-shaped beams, so that, when the spacing of longitudinal stiffeners decreases, flexural capacity tends to increase, especially in the elastic buckling zone.