Due to the negative environmental impact of cement production, partial replacement of cement with by-products and wastes has been drawn much attention. However, the reduction in binder fraction can affect the fresh and mechanical properties of cement-based materials. In this study, the effects of nano and micro Fe3O4 on the fresh and mechanical properties of plain mortar as well as mortar modified with fly ash were investigated. The preliminary study shows that the optimum dosage of Fe3O4 particle in mortar is 5% nano Fe3O4 (NF). Further study was conducted with 5% NF where cement was replaced by up to 30% fly ash. The results showed that the partial replacement of binder with fly ash at 10% decreases the compressive strength from 18.8 MPa to 15.4 MPa at 28 days, and the least compressive strength was achieved in the presence of 30% fly ash (10.3 MPa). Despite an insignificant change in splitting tensile strength in the presence of 10% fly ash (2.27 MPa), further increase in fly ash content considerably decreased the splitting tensile strength. Although the increasing dosage of fly ash decreased the strength of mortar, the presence of NF could counteract these effects and improve the strength development.