This study explored the effects of a professional development (PD) on neuroeducation general pedagogical knowledge and instructional strategies for three cohorts of in-service teachers on their teaching self-efficacy beliefs. Teacher self-efficacy is one of the most influential factors for teaching practices, student outcomes, and teacher persistence in the profession. We utilized a neuroeducation instructional framework in the PD to provide knowledge of learning sciences research to inform practices across all curricula. The PD was designed to help teachers translate research into general pedagogical practices to address topics such as classroom emotional climate, student attention and memory, meaningful application of knowledge, and creative problem solving. Two cohorts of teachers engaged in the PD for five months during two consecutive spring semesters, and one cohort participated in a two-week session of the PD. We used a pretest-posttest quasi-experimental design and findings revealed increases in teachers' self-efficacy beliefs in all cohorts. Implications for practice and research are discussed.