Nonlinear gyrokinetic simulations of impurity transport are compared to experimental impurity transport for the first time. The GYRO code (Candy and Waltz 2003 J. Comput. Phys. 186 545) was used to perform global, nonlinear gyrokinetic simulations of impurity transport for a standard Alcator C-Mod, L-mode discharge. The laser blow-off technique was combined with soft x-ray measurements of a single charge state of calcium to provide time-evolving profiles of this non-intrinsic, non-recycling impurity over a radial range of 0.0 ⩽ r/a ⩽ 0.6. Experimental transport coefficient profiles and their uncertainties were extracted from the measurements using the impurity transport code STRAHL and rigorous Monte Carlo error analysis. To best assess the agreement of gyrokinetic simulations with the experimental profiles, the sensitivity of the GYRO predicted impurity transport to a wide range of turbulence-relevant plasma parameters was investigated. A direct comparison of nonlinear gyrokinetic simulation and experiment is presented with an in depth discussion of error sources and a new data analysis methodology.