Background: Acute mountain sickness (AMS) is the most common type of high-altitude sickness. The incidence of AMS varies by mountain location, trail characteristics, and study design. The lack of local epidemiology data has driven us to investigate the incidence and severity of AMS and its associated factors at Mount Kinabalu, Malaysia. Methods: A cohort study was conducted to collect data from climbers after days 1 (3272 m) and 2 (4095 m) of ascent. A self-administered questionnaire was used to explore climbers' demographic and climb characteristics, history of AMS, alcohol exposure, and AMS prevention measures. The Lake Louis score 2018 was used to assess the presence and severity of AMS (cutoff ≥3). Univariate and multivariable logistic regressions were performed to determine the factors associated with the development of AMS on day 2. Results: Data from 345 climbers were analyzed. The incidence of AMS was 23.9% (95% confidence interval [CI] 19.5%-28.7%) and 21.7% (95% CI 17.5%-26.3%) on days 1 and 2, respectively. The majority were mild cases. Experiencing AMS on day 1 (odds ratio [OR] = 12.88; 95% CI 6.71-24.75), alcohol consumption (OR = 3.73; 95% CI 1.66-8.39), receiving guide advice on day 1 (OR = 0.49; 95% CI 0.26-0.93), and age (OR = 0.96; 95% CI 0.93-0.99) were significant determinants of AMS at Mount Kinabalu. Gender, history of AMS, past exposure to high altitude, ascending time, water intake, acetazolamide use, physical fitness, pulse rate, and peripheral capillary oxygen saturation (SpO2) were not associated with AMS at Mount Kinabalu. Conclusion: Future analysis with age strata is required to ascertain the association of age with AMS. Our research has signposted a strong call for collaborative efforts to improve the provision of hiking advice and discourage alcohol sales to mitigate the risk of AMS among Mount Kinabalu climbers.