Abstract For designing low-impedance magnetic tunnel junctions (MTJs), it has been found that tunneling magnetoresistance (TMR) strongly correlates with the insulating barrier thickness, imposing a fundamental question on the relationship between spin polarization of ferromagnet and the insulating barrier thickness in MTJs. Here, we investigate the influence of alumina barrier thickness on tunneling spin polarization (TSP) through a combination of theoretical calculations and experimental verification. Our simulating results reveal a significant impact of barrier thickness on TSP, exhibiting an oscillating decay of TSP with the barrier layer thinning. Experimental verification is realized on FeNi/AlO$_x$/Al superconducting tunnel junctions to directly probe the spin polarization of FeNi ferromagnet using Zeeman-split tunneling spectroscopy technique. These findings provide valuable insights for the design of high-performance spintronic devices, particularly in applications such as MRAM, where precise control over the insulating barrier layer is crucial.