Existing research has shown that emotional intelligence (EI) and teacher self-efficacy (TSE) play an important role in the work of in-service teachers. However, there is limited research on these variables and their associations among pre-service teachers. Also, the cultural context is expected to influence EI and TSE. Therefore, this study examined the associations between EI and TSE in two cultures, Finland and Greece. A comprehensive evaluation of EI was done by including both trait EI and ability EI measures. Data from primary education student teachers from Finland (N = 82) and Greece (N = 117) were collected online. The measures of EI and TSE exhibited full configural and metric measurement invariance and partial scalar and residual/strict measurement invariance across cultures. Finnish students had significantly higher scores on all EI variables than Greek students, whereas Greek students scored higher on self-efficacy for student engagement. Structural equation modelling showed that trait EI was moderately associated with one facet of ability EI, namely emotional management, but not with emotional understanding, implying that trait EI and ability EI can be considered as partially distinct constructs. A statistically significant positive association between trait EI and TSE was found in Finland, whereas a statistically significant negative association between ability EI and TSE was found in Greece. The findings have important implications for the models of trait and ability EI, for understanding the links between trait and ability EI and TSE during the critical phase of teacher education in the two cultural contexts studied, and for culturally informed teacher education.