Overwhelming evidence demonstrating the benefits of active-learning pedagogy has led to a shift in teaching that requires students to interact more in the classroom. To date, few studies have assessed whether there are gender-specific differences in participation in active-learning science, technology, engineering, and mathematics (STEM) courses, and fewer have looked across different types of classroom participation. Over two semesters, we observed an introductory biology course at a large research-intensive university and categorized student participation into seven distinct categories to identify gender gaps in participation. Additionally, we collected student grades and administered a postcourse survey that gauged student scientific self-efficacy and salience of gender identity. We found that men participated more than expected based on the class composition in most participation categories. In particular, men were strongly over-represented in voluntary responses after small-group discussions across both semesters. Women in the course reported lower scientific self-efficacy and greater salience of gender identity. Our results suggest that active learning in itself is not a panacea for STEM equity; rather, to maximize the benefits of active-learning pedagogy, instructors should make a concerted effort to use teaching strategies that are inclusive and encourage equitable participation by all students.