Conventional measurement methods for above-ground biomass (AGB) are time-consuming, inaccurate, and labor-intensive. Unmanned aerial systems (UASs) have emerged as a promising solution, but a standardized procedure for UAS-based AGB estimation is lacking. This study reviews recent findings (2018–2022) on UAS applications for AGB estimation and develops a vegetation type-specific standard protocol. Analysis of 211 papers reveals the prevalence of rotary-wing UASs, especially quadcopters, in agricultural fields. Sensor selection varies by vegetation type, with LIDAR and RGB sensors in forests, and RGB, multispectral, and hyperspectral sensors in agricultural and grass fields. Flight altitudes and speeds depend on vegetation characteristics and sensor types, varying among crop groups. Ground control points (GCPs) needed for accurate AGB estimation differ based on vegetation type and topographic complexity. Optimal data collection during solar noon enhances accuracy, considering image quality, solar energy availability, and reduced atmospheric effects. Vegetation indices significantly affect AGB estimation in vertically growing crops, while their influence is comparatively less in forests, grasses, and horizontally growing crops. Plant height metrics differ across vegetation groups, with maximum height in forests and vertically growing crops, and central tendency metrics in grasses and horizontally growing crops. Linear regression and machine learning models perform similarly in forests, with machine learning outperforming in grasses; both yield comparable results for horizontally and vertically growing crops. Challenges include sensor limitations, environmental conditions, reflectance mixture, canopy complexity, water, cloud cover, dew, phenology, image artifacts, legal restrictions, computing power, battery capacity, optical saturation, and GPS errors. Addressing these requires careful sensor selection, timing, image processing, compliance with regulations, and overcoming technical limitations. Insights and guidelines provided enhance the precision and efficiency of UAS-based AGB estimation. Understanding vegetation requirements aids informed decisions on platform selection, sensor choice, flight parameters, and modeling approaches across different ecosystems. This study bridges the gap by providing a standardized protocol, facilitating widespread adoption of UAS technology for AGB estimation.