Objective: To describe the knowledge, attitudes, and practices (KAPs) profile on bacterial resistance and antibiotic in the general population of Medellín. Material and methods: A cross-sectional study was conducted from September to December 2022, with 902 participants selected through stratified sampling with proportional allocation of different sectors of the city. The KAP scale was developed through a literature review, elimination of duplicate items, validity assessment, and reliability evaluation using Cronbach’s alpha. Each item was presented with absolute and relative frequencies on a Likert scale, with a total score ranging from 0 to 100, where a higher score indicates better knowledge, attitudes, and practices. Comparisons were made using Mann-Whitney U, Kruskal-Wallis H, and linear regression. Results: The knowledge score median was 73.3 (IQR 63.3–93.3), with 36.9% reporting that antibiotics can be stopped once symptoms improve and 26.1% considering them as analgesics or antipyretics. The attitudes score was 83.3 (IQR 73.3–93.3), with 95.3% expressing concern about the impact on their health or that of their family and over 90% agreeing that more information is needed on antibiotic resistance. The practice score was the lowest at 63.9 (IQR 50–75), with 48% having been prescribed antibiotics at the pharmacy and 42.6% taking them to treat flu symptoms. Economic status (β 2.645), education in health-related areas (β 6.224), gender (β 2.892), and education level (β 3.257) determined knowledge. Knowledge (β 0.387), gender (β 2.807), and education level (β 0.686) influenced attitudes, but practices were only determined by Knowledge (β 0.084) attitudes (β 0.552) and age group (β 2.858). Conclusions: Knowledge about antibiotics and bacterial resistance does not significantly influence the practices of the population. Therefore, interventions aimed at improving knowledge need to be reconsidered as they may not contribute to the appropriate use of antibiotics and prevention of resistance to these drugs.