Nighttime light data can characterize urbanization, economic development, population density, energy consumption and other human activities. Additionally, carbon dioxide (CO2) emissions are closely related to the scope and intensity of human activities. In this study, we assess the utility of nighttime light data as a powerful tool to reflect CO2 emissions from energy consumption, analyze the uncertainty associated with different nighttime light data for modeling CO2 emissions, and provide guidance and a reference for modeling CO2 emissions based on nighttime light data. In this paper, Mainland China was taken as a case study, and nighttime light datasets (the Defense Meteorological Satellite Program’s Operational Linescan System (DMSP-OLS) nighttime light data and the Suomi National Polar-Orbiting Partnership Visible Infrared Imaging Radiometer Suite (NPP-VIIRS) nighttime light data) as well as a global gridded CO2 emissions dataset (PKU-CO2) were used to perform simple regressions at provincial, prefectural and 0.1° × 0.1° grid levels, respectively. The analyses are aimed at exploring the accuracy and uncertainty of DMSP-OLS and NPP-VIIRS nighttime light data in modeling CO2 emissions at different spatial scales. The improvement of nighttime light index and the potential factors influencing the effects of modeling CO2 emissions based on nighttime light datasets were also explored. The results show that DMSP-OLS is superior to NPP-VIIRS in modeling CO2 emissions at all spatial scales, and the bigger the scale, the more evident the advantages of DMSP-OLS. When modeling CO2 emissions with nighttime light datasets, not only the total amount of lights within a given statistical unit but also the agglomeration degree of lights should be taken into account. Furthermore, the geographical location and socio-economic conditions at the study site, such as gross regional product per capita (GRP per capita), population, and urbanization were shown to have an impact on the regression effect of the nighttime lights-CO2 emissions model. The regression effect was found to be better at higher latitude and longitude areas with higher GRP per capita and higher urbanization, while population showed little effect on the regression effect of the nighttime lights - CO2 emissions model. The limitation of this study is that the thresholds of potential factors are unclear and the quantitative guidance is insufficient.