Background: Antibiotic administration can induce a life-threatening condition known as anaphylaxis, so correct administering antibiotics and efficient management of anaphylaxis from nurses is crucial for lifesaving all patients. Aim: Assess the effect of nursing educational protocol on the nurses' knowledge and practice regarding antibiotic administration and anaphylaxis management. Method: Aquasi-experimental research design (pre- and post-test). Setting: The study was conducted in the medical and pediatric departments at Assuit University Hospital. Subjects: 300 nurses employed in the medical and pediatric departments. Tools: (1)a structured questionnaire sheet to assess nurses` knowledge about antibiotic administration and anaphylaxis management (2) Pre/ Post Nurses' practice intradermal (ID ) hypersensitivity skin test. Results: The mean age of the studied nurses was 36.1±7.0 years,82.0% of them were female , and highly statistically significant differences, indicating improvement was founded between nurses` knowledge and practice before and after implementing nursing educational protocol about antibiotic administration and anaphylaxis management (P=0.001***). Conclusion: The application of nursing educational protocols was effective and successfully improved the nurses' knowledge and practice regarding antibiotic administration and anaphylaxis management. Recommendation: Establishing nursing educational protocols in hospitals will improve the nurses' knowledge and practice about how to manage anaphylaxis occurrences.