MODIS has 20 reflective solar bands (RSB) and 16 thermal emissive bands (TEB). Compared to its heritage sensors, MODIS was developed with very stringent calibration uncertainty requirements. As a result, MODIS was designed and built with a set of on-board calibrators (OBC), which allow key sensor performance parameters and on-orbit calibration coefficients to be monitored and updated. In terms of its calibration traceability, MODIS RSB calibration is reflectance based using an on-board solar diffuser (SD) and the TEB calibration is radiance based using an on-board blackbody (BB). In addition to on-orbit calibration coefficients derived from its OBC, calibration parameters determined from sensor pre-launch calibration and characterization are used in both the RSB and TEB calibration and retrieval algorithms. This paper provides a brief description of MODIS calibration methodologies and an in-depth analysis of its on-orbit calibration uncertainties. Also discussed in this paper are uncertainty contributions from individual components and differences due to Terra and Aqua MODIS instrument characteristics and on-orbit performance.