The goal of this study was to compare the accuracy of kinematic measurements obtained using the 2-D video-based kinematic motion analysis (KMA) software Kinovea (version 0.9.5; http://www.kinovea.org) with 3-D KMA in healthy dogs. In this prospective study, 3-D marker-based KMA (VICON-Nexus, version 2.12.1, and Procalc, version 1.6; VICON Motion Systems Ltd) was performed on healthy dogs (body weight ≥ 20 kg; height at withers > 50 cm) walking on a treadmill (study period: November 2022). Simultaneously, dogs were video recorded by 1 smartphone (iPhone SE; Apple Inc) at a 1.50-m distance perpendicular to the shoulder (60 frames per second; 1,920 X 1,080 pixels) for KMA using Kinovea. Joint angle and joint angle velocity of the shoulder, elbow, carpus, hip, stifle, and tarsus were calculated for 6 synchronized gait cycles. Each gait cycle was divided into 10 increments. The difference between 3-D KMA and Kinovea was assessed for each parameter using robust linear mixed-effects models. 34 dogs were included. The estimated joint angle difference between 3-D KMA and Kinovea was less than 2° for all shoulder and elbow gait cycle increments. For the carpus, hip, stifle, and tarsus, the difference was less than 2° in 9, 5, 4, and 4 out of 10 gait cycle increments, respectively. Kinovea provides accurate kinematic data for the shoulder and elbow of healthy dogs. Carpal, hip, stifle, and tarsal kinematics were less accurate. The use of Kinovea for clinical and research purposes remains limited. Future Kinovea-based studies are needed to investigate the accuracy of carpal, hip, stifle, and tarsal kinematics.