Isoenzymes are variant forms of the same enzyme activity that exhibits different tissue distribution. When cells are damaged, intracellular content leaks into the blood stream, making them potential biochemical markers. Undergraduate health sciences students are expected to interpret laboratory findings of serum levels of biochemical markers and understand their relevance for diagnosis, prognosis and treatment. This study aimed to design an electrophoresis laboratory class that challenges health sciences students to discuss the experimental approach and interpret the results obtained from electrophoresis of Creatine Kinase (CK) isoenzymes separation. Mixtures of dyes were used to simulate the serum composition of CK isoenzymes in different clinical conditions and the resulting bands were quantified by densitometry. Two online questionnaires assessed students' learning progress and acceptance of the activity. Undergraduate students from biomedicine (n=10) and medicine (n=42) programs participated in a 120-minute pilot class at Universidade Federal Fluminense, Brazil. The post-class test showed a 21.9% increase in correct answers compared to the pre-class test. In the survey, students positively rated the laboratory class, expressing improved understanding of theoretical concepts, recognizing the importance of basic biochemistry for their future careers and comprehending the electrophoretic evaluation of CK isoenzymes.