Antibiotics have saved millions of lives and changed the history of infectious diseases. However, in recent years, antibiotic resistance has become a major global public health problem. Likewise, an increasing trend of antibiotic resistance (ABR) is observed in Malaysia. This study aims to determine the knowledge, attitude and practice (KAP) towards antibiotics use and its resistance among the general public in Klang Valley, Malaysia. Our hypothesis is the general public has a low level of KAP towards antibiotic use and its resistance. A cross-sectional study was distributed through a pre-validated questionnaire to all Klang Valley residents who are ≥18 years old through email and social media. Vulnerable residents and those from non-English speaking backgrounds were excluded. All data were analysed by SPSS software version 27.0. A total of 408 participants were recruited. Based on the scoring system of the study it was found that 47.0% had moderate knowledge, 41.7% had good attitude and 59.6% had good practice towards antibiotic use and its resistance. Overall, 53.4% recruited participants had moderate KAP towards antibiotic use and its resistance. Chi-Square test of independence was used to determine the association of socio-demographic variables and KAP towards antibiotic use and its resistance. The results indicated that only one categorical variable “area” was statistically significant with a p value of 0.041. The mean± SD values indicated that those individuals living in Selangor have a comparatively higher KAP towards antibiotic use and its resistance as compared to those living in Kuala Lumpur (KL) most probably due to higher educational levels. This study provided us with baseline evidence about the KAP regarding antibiotic use among the general public in Malaysia. Hence, it will be useful in guiding further interventions to improve awareness about antibiotics use and enhance antimicrobial stewardship in our community.