AbstractGrassland aboveground biomass (AGB) is a key variable to measure grassland productivity, and accurate assessment of AGB is important for optimizing grassland resource management and understanding carbon, water, and energy fluxes. Current approaches on large scales such as the Mongolian Steppe Ecosystem often combine field measurements with optical and/or synthetic aperture radar (SAR) data. Meanwhile, especially the representativeness of the field measurements for large‐scale analysis have seldom been accounted for. Therefore, we provide the first remotely sensed AGB product for central and Eastern Mongolia which (1) uses random forest (RF), (2) is fully validated against over 600 field samples, and (3) applies a novel method, dissimilarity index (DI), to derive the area of applicability of the model with respect to the training data. Therefore, different remote sensing data sources such as multi‐scale and multi‐temporal optical images—Worldview 2 (WV2), Sentinel 2 (S2), and Landsat 8 (L8) in combination with SAR data are tested for their suitability to provide an area‐wide estimation on large scale. The results showed that the AGB prediction by combining Sentinel 1 (S1) and S2 using RF had the highest accuracy. Furthermore, the model was applicable to at least 72.61% of the steppe area. Areas where the model was not applicable are mostly distributed along the edges of grassland. This study demonstrates the potential of combining Sentinel‐derived indices and machine learning to provide a reliable AGB prediction for grassland for extremely large ecosystems with strong climatic gradients.