Abstract Introduction Theory and data suggest that attentional bias (AB) to drug-related cues should be associated with craving when smoking motivation is high, and that AB should be predictive of drug use when immediate use is possible. The current study is the first to test these propositions in smokers in a controlled laboratory environment. Aims and Methods Ninety daily smokers were randomly assigned to a high smoking motivation (nicotine-deprived and/or smoking cue exposure) or low smoking motivation (non-deprived and/or control cue exposure) condition. Participants engaged in an AB task in which they viewed smoking and matched control pictures while their eye movements were continuously monitored. Participants were then given the option to smoke, and latency to first puff and number of puffs were coded. Results High motivation smokers had significantly higher urges to smoke (p < .001) and shorter latencies to smoke (p = .001) than low motivation smokers, but AB measures (ie, dwell time and initial fixation bias scores) and number of puffs did not differ across groups (ps > .45). As predicted, the association between dwell time bias scores and urge to smoke was stronger in the high (r = .47) than low (r = .18) smoking motivation condition, but this difference failed to reach significance (p = .068). Contrary to predictions, neither AB measure was significantly associated with smoking behavior (SB). Internal reliability was excellent for dwell time bias scores (alpha = .90) but very low for initial fixation bias scores (alpha = .20). Conclusions Maintenance of attention on drug-related cues may be a valid index of incentive motivation. Importantly, however, these dwell time bias scores were not predictive of actual SB. Implications This study tested key predictions made by theoretical accounts of addiction that emphasize AB to drug-related cues as fundamental components of the development and maintenance of drug use. Namely, this is the first experimental study in smokers to test whether AB to smoking-related cues is associated with craving when smoking motivation is high and whether AB predicts SB assessed immediately after the AB task. As predicted, the association between AB and craving was stronger in smokers randomly assigned to a high rather than a low smoking motivation condition. Contrary to predictions, AB did not predict SB.