Background: Electronic Portfolio is a learning support tool that emphasizes learning process, experience and professional development to present the achievement. Motivation to use electronic portfolio is needed to energize the desire to learn. Aim: the present study aimed to assess nursing teaching staff and students' readiness about using electronic portfolio and its relation to students’ academic motivation. Research design: A descriptive correlational design was utilized. Setting: This study was conducted at the Faculty of Nursing, Benha University. Subjects: All available of nursing teaching staff 112 and stratified random sample consisted of 1804 of nursing students. Tools of data collection: Two tools were used to collect data; I) Electronic Portfolio readiness questionnaire, II) Nursing students academic motivation questionnaire. Results: showed that more than three fifths of nursing teaching staff had high readiness level about using electronic portfolio, also, showed that nearly two thirds of nursing students had high level of readiness about using electronic portfolio. And, the majority of nursing students had high academic motivation level. Conclusion: there was a statistical significant correlation between total nursing teaching staff readiness toward using electronic portfolio and total academic motivation among nursing students. Recommendations: conducting workshops for nursing teaching staff & nursing students about how to apply electronic portfolio and using electronic portfolio as a method for assessment.