As a specific force sensor, the tri-axis accelerometer is one of the core instruments in an inertial navigation system (INS). During navigation, its measurement error directly induces constant or alternating navigation errors of the same order of magnitude. Moreover, it also affects the estimation accuracy of gyro drift coefficients during the initial alignment and calibration, which will indirectly result in navigation errors accumulating over time. Calibration can effectively improve measurement accuracy of the accelerometer. Device-level calibration can identify all of the parameters in the error model, and the system-level calibration can accurately estimate part of these parameters. Combining the advantages of both the methods and making full use of the precise angulation of the space-stabilized platform, this paper proposes a three-stage accelerometer self-calibration technique that can be implemented directly in the space-stable INS. The device-level calibration is divided into two steps considering the large amount of parameters. The first step is coarse calibration, which identifies parameters except for the nonlinear terms, and the second step is fine calibration, which not only identifies the nonlinear parameters, but also improves the accuracy of the parameters identified in the first step. The follow-on system-level calibration is carried out on part of the parameters using specific force error and attitude error to further improve the calibration accuracy. Simulation result shows that by using the proposed three-stage calibration technique in the space-stable INS, the estimation accuracy of accelerometer error can reach 1×10−6 normalg order of magnitude. Experiment results show that after the three-stage calibration, the accuracy of latitude, longitude, and attitude angles has increased by over 45% and the accuracy of velocity has increased by over 22% during navigation.