Bolted connections are crucial in joining mechanical assemblies and ensuring the integrity and reliability of structural components. This study proposes a method for estimating bolt loosening life by practically applying material fatigue life prediction methods. First, the study employed the linear cumulative damage rule to predict the loosening life of single bolts under two-block loading conditions. Second, a test device with two bolt attachment points on a single structure was fabricated to model the multi-bolted structure, and tests were conducted. Finite element analysis (FEA) analysis was employed to identify vulnerable points. The loosening life of single bolts predicted using the linear cumulative damage rule exhibited enhanced accuracy within a ±1.2× error band compared with the experimental data despite variations in bolt types and test conditions. The FEA results for the multi-bolt structure demonstrated that the loosening life could be predicted by identifying vulnerable points and estimating the displacements. This study effectively predicts the bolt loosening life, offering valuable data for the reliability assessment of bolted structures.