Objective: The aim of the study was to assess the knowledge, attitudes and practices of students regarding the use of antibiotics in Punjab, Pakistan. Participants: 525 medical and non-medical students from Punjab in Pakistan. Methods: The t-test and ANOVA were used to compare the average response of respondents. Chi-square test was used to measure the association of different elements. Results: The mean age was 20.78 ± 2.10%. About 14% of the students agreed about the appropriateness of antibiotics for viral infections, and 15% of students said they stopped taking the drugs when symptoms subsided. 65.7% of students took antibiotics only when prescribed by a doctor and 54% bought antibiotics without prescription. Statistically significant results were found among the students who had heard about the antibiotic resistance (p < 0.05). Conclusion: This study will help assess the adequacy of current educational campaigns, maximize rationalization of antibiotic use, and minimize gaps in knowledge and attitudes.