Inwardly rectifying potassium (Kir) channels modulate cellular excitability, membrane potential, and secretion of neurotransmitters and hormones. Kir channels with the strongest inward rectification belong to the Kir2 family. In this report, polyclonal monospecific affinity-purified antibodies against the less conserved carboxy-terminal sequences of Kir2.1, Kir2.2, Kir2.3, and Kir2.4 were used to analyze the detailed distribution of all members of the Kir2 family in the rat central nervous system. Kir2 channel expression is detected in neurons but not in glial cells. Kir2 protein distribution confirms the basic mRNA localization pattern given by in situ hybridization. Kir2.1 is detected throughout the whole brain but in particular subsets of neurons with highest expression in olfactory bulb and superior colliculus. Kir2.2 immunoreactivity is primarily displayed in several forebrain nuclei, hypothalamus, cerebellum, and spinal cord. The Kir2.3 subunit is predominantly localized in olfactory bulb, basal ganglia, cortex, and cerebellar Purkinje cells. In contrast, Kir2.4-positive staining is detected at significantly lower levels in most neurons throughout the rat brain with highest expression in brainstem motoneurons. Thus, our data show a more widespread distribution of Kir2.4 than previously determined. In summary, the widespread presence of all four Kir2 channel subunits in the rat brain provides further evidence for their important role in central signal processing and neural transmission.