Introduction: Widespread use of chemotherapeutic drugs in the treatment of cancer can lead to higher health hazards among nurses who handle and administer such drugs, so they should know how to protect themselves from the effects of chemotherapy. Aim of study: was to evaluate the effect of implementing Safety Handling Protocol for Chemotherapy (SHPC) on nurses' knowledge and practical skills. Research design: A Quasi – experimental research design was utilized to carry out this study. Setting: this study was conducted at Minia Oncology Center. Subjects: all available nurses (50) who are working in outpatient and inpatient chemotherapy department. Tools: three tools were designed by researcher and utilized; Arabic administered questionnaire, structured observational checklist and safe handling chemotherapy protocol (booklet). Results: nurses had poor knowledge regarding safe handling chemotherapy. The mean score of total nurses knowledge statistically improved from (12.9± 4.8 to 25.4 ± 2.0 & 22.7 ± 2.8 respectively) in pre, immediately, and after 2 months of implementing (SHPC) with p value (.0001**). Nearly all nurses have unsatisfactory practical skill level before implementing (SHPC) which significantly changed after implementing it (90%, 18%, 20% respectively) with p value (.0001**). Conclusion: the results of this study revealed significant improvement of nurses' knowledge, and practical skills among the nurses handling chemotherapeutic drugs after implementing (SHPC). Recommendations: health care professionals who are responsible for handling chemotherapeutic drugs should be adherent to implementation of (SHPC).