Bamboo is susceptible to longitudinal splitting, which is mainly driven by its shear properties parallel to the fibers. In this work, bowtie tests including 124 Moso bamboo internode and node specimens were conducted to investigate the shear behaviors of Moso bamboo in shear parallel to the fibers. Based on the test results, the difference in shear strength between internode and node specimens using the analysis of variance (ANOVA) was quantitatively evaluated. The distribution of shear strength was fitted using Normal, Log-normal, and Weibull distributions, and the corresponding standard value was calculated using the non-parametric method. The design value was obtained based on reliability analysis. Additionally, prediction formulas for shear strength were fitted using univariate and multiple regression analyses based on outer diameter, wall thickness, and density. Various shear stress-strain constitutive models were established and validated. The research results indicated that there is no statistically significant difference in shear strength between internode and node specimens with the value of 17.17 MPa and 17.48 MPa, respectively. The distribution of shear strength followed the Normal and Log-normal distributions, but not the Weibull distribution. The standard value and design value of shear strength were 14.91 and 9.47 MPa, respectively. The shear strength was found to be negatively correlated with the outer diameter and wall thickness, but positively correlated with the density. The recommended strength prediction formulas and stress-strain constitutive models match well with the test results. This study can serve as a reference for the improvement of bamboo standard and design basis of Moso bamboo.