Insulator-based dielectrophoresis (iDEP) has been explored as a powerful analytical technique in recent years. Unlike with larger entities such as cells, bacteria or organelles, the mechanism of iDEP transport of proteins remains little explored. In this work, we extended the pool of proteins investigated with iDEP in nanostructured devices with β-galactosidase. Our work indicates that β-galactosidase shows concentration due to negative DEP which we compare to DEP response of immunoglobulin G (IgG) encapsulated in micelles also showing negative DEP. Experimental observations are further compared with numerical simulations to elucidate the influence of electrokinetic transport and the magnitude of DEP mobility. Numerical simulations suggest that the DEP mobility calculated using the classical model underestimates the actual contribution of DEP on the experimentally monitored concentration effect of proteins. Moreover, we observed a unique voltage dependent β-galactosidase concentration which we attribute to an additional factor influencing the protein concentration at the nanoconstrictions, namely ion concentration polarization. Our work aids in understanding factors influencing protein iDEP transport which is required for the future development of protein preconcentration or separation methods based on iDEP.