Cellular sensors and their signalling components form the first step in the recognition of intracellular pathogens by orchestrating innate immune activation. Many of these proteins undergo subcellular relocalisation upon activation, including key regulators of immune and inflammatory responses. Identifying novel proteins that translocate in response to immune stimuli can reveal new mechanisms governing innate immunity and antiviral defence. We have developed a spatial proteomics workflow combining subcellular fractionation with quantitative mass spectrometry to globally profile protein translocation following type I interferon (IFN) stimulation. Known mediators of the IFN response, IRF9 and STAT1/2, exhibited cytoplasmic to nuclear translocation, validating our approach. We identified 25 proteins that reproducibly translocated from the cytoplasm to the nucleus. One of these was a previously uncharacterised protein, which we have named INCH (Innate immune nucleocytoplasmic checkpoint), which exhibited striking nucleocytoplasmic translocation but did not exhibit interferon induction that would be typical of an interferon stimulated gene. Immunoprecipitation followed by mass spectrometry revealed that INCH interacts predominantly with ribosomal and double stranded RNA binding proteins. Depletion of INCH resulted in significantly elevated basal ISG expression and IRF3 activation in specific cell types. These findings indicate that INCH may act as a selective cellular checkpoint in innate immune signalling. Overall, by integrating advanced proteomics with cell biology, we provide a comprehensive subcellular map of the basal and IFN stimulated response across different cell types and have identified INCH as a novel, cell type-specific IFN responsive regulator of innate immunity.