For the teaching profession, prosociality is a relevant skill. From this perspective, the present study aimed to examine the psychometric properties of the Adult Prosocialness Behavior Scale (APBS) in a professional setting of primary school Chilean teachers (n = 1404; mean age = 41.4; SD = 10.8; 77.4% women). ESEM and CFA were applied to evaluate its factor structure, internal consistency, factorial reliability, and invariance. The results supported a bifactor ESEM model in which prosociality was represented by a general latent factor and four specific factors (helping, sharing, caring, and empathy). A predominance of the unidimensional component of the APBS was observed (general factor, ECV = 0.938; ωh = 0.945), with adequate reliability (α = 0.932; ω of the factor general = 0.968) and invariance of the measurement in terms of gender. Its adequacy was confirmed by a good level of fit (RMSEA = 0.042 90%CI [0.036–0.049]; SRMR = 0.012; CFI = 0.995; TLI = 0.988). It is concluded that the APBS is a suitable instrument to measure prosociality in the professional setting studied. Its general and specific dimensions are relevant to the prosocial behaviors currently required of teachers. Future studies could contribute evidence of the transcultural validation of the APBS in other educational contexts in order to undertake comparative studies.