Accurate mapping of above-ground biomass (AGB) is essential for carbon stock quantification and climate change impact assessment, particularly in mountainous areas. This study applies a random forest (RF) regression model to predict the spatial distribution of AGB in Usho (site A) and Utror (site B) forests located in the northern mountainous region of Pakistan. The predicted maps elucidate AGB variations across these sites, with non-forest areas excluded based on an normalized difference vegetation index (NDVI) threshold value of <0.4. Three different combinations of input datasets were used to predict the biomass, including spectral bands (SBs) only, vegetation indexes (VIs) only, and a combination of both spectral bands and vegetation indexes (SBVIs). Utilizing SBs, the biomass ranged between 150 and 286 mg/ha in site A and 99 and 376 mg/ha in site B. Meanwhile, using VIs indicated a biomass range of 163 Mg/ha–337 Mg/ha and 131–392 Mg/ha for sites A and B, respectively. The combination of spectral bands and vegetation indexes yielded AGB values of 145–290 Mg/ha in site A and 116–389 Mg/ha in site B. The northern and western regions of site A, characterized by higher altitudes and lower forest density, notably showed lower biomass values than other regions. Conversely, similar regions in site B, situated at lower latitudes, demonstrated different biomass ranges. The RF model exhibited robust accuracy, with R2 values of 0.74 and 0.83 for spectral bands and vegetation indexes, respectively. However, with a combination of both, an R2 of 0.79 was achieved. Furthermore, altitudinal gradients significantly influence the biomass distribution across both sites, with specific elevation ranges yielding optimal results. The AGB variation along the slope further corroborated these findings. In both sites, the western aspects showed the highest biomass across all combinations of input datasets. The variable importance analysis highlighted that ARVI8a, NDI45, Band12, Band11, TSAVI8, and ARVI8a are significant predictors in sites A and B. This comprehensive analysis enhances our understanding of AGB distribution in the mountainous forests of Pakistan, offering valuable insights for forest management and ecological studies.