Assessing stream fish habitat associations across contrasting ecosystems can inform generality of habitat predictions. We tracked Cottus carolinae (Banded Sculpin) in Little Creek, TN, to test transferability of habitat predictions developed from independent studies. Predictions included shifting habitat use across size classes (prediction 1), over the diel period (prediction 2), and during variable flows (prediction 3), as well as maintaining associations with depth, velocity, and substrate gradients across scales (prediction 4). Size 1 (80–99 mm TL) and size 2 (100–140 mm TL) Banded Sculpin used similar habitats (prediction 1 not supported), shifted to pools with little cover at night (prediction 2 supported), and adjusted habitat uses according to flow (prediction 3 supported), and depth, velocity, and substrate associations were similar for small and large streams when size classes were combined (prediction 4 supported). Our synthesis highlights consistencies in fish habitat associations that manifest due to behavioral, morphological, and physiological constraints that operate across ecosystems.