Radiology is a vital diagnostic tool for multiple disorders that plays an essential role in the healthcare sector. Nurses are majorly involved in a healthcare setting by accompanying patients during the examination. Thus, nurses tend to be exposed during inward X-ray examination, requiring them to keep up with radiation use safety. However, nurses’ competence in radiation is still a concept that has not been well studied in Malaysia. The study aimed to define the level of usage understanding and radiation protection among Malaysian nurses. In this research, a cross-sectional survey was conducted among 395 nurses working in hospitals, clinics, and other healthcare sectors in Malaysia. The survey is based on the developed Healthcare Professional Knowledge of Radiation Protection (HPKRP) scale, distributed via the online Google Forms. SPSS version 25.0 (IBM Corporation) was used to analyze the data in this study. Malaysian nurses reported the highest knowledge level in radiation protection with a mean of 6.03 ± 2.59. The second highest is safe ionizing radiation guidelines with 5.83 ± 2.77, but low knowledge levels in radiation physics and radiation usage principle (4.69 ± 2.49). Therefore, healthcare facilities should strengthen the training standards for all nurses working with or exposed to radiation.