In the present work, nickel-doped iron oxide (NixFe3−xO4) nanoparticles with different concentration of nickel (x = 0, 0.05, 0.1, and 0.15) have been prepared by co-precipitation method. These prepared nanoparticles have been characterized by using x-ray diffractometer, thermo gravimetric analysis and differential scanning calorimetry, Fourier transform infrared spectroscopy, scanning electron microscopy, vibrating sample magnetometer, and UV-Visible spectroscopy to study their structural, thermal, morphological, magnetic, and optical properties, respectively. The x-ray diffraction confirms the formation of single-phase inverse spinel cubic structure of NiFe3O4 nanoparticles. Crystallite size has been estimated by the full width at half maximum of the most intense x-ray diffraction peak where vibrational and stretching modes of metal-oxygen bonds in 872 cm are shown in Fourier transform infrared spectra which confirms the formation of nanoparticles. The thermal analysis revealed that the transition temperature and stability increases with increasing Ni concentration. The surface morphology indicated that the particles are spherical in shape with some agglomeration. The magnetic measurement revealed that the coercivity and anisotropy increases with nickel doping in magnetite nanoparticles. The optical analysis revealed that direct and indirect both types of band gap increases when the particle size decreases because the absorption spectra shift toward smaller wavelength. The blue shift confirms the formation of nanoparticles.