AbstractLiver samples from female and male mice of two subspecies, Mus musculus musculus and Mus musculus domesticus, were investigated by a combination of 2‐DE and MALDI‐MS. The image analysis of the generated 2‐DE patterns revealed several protein spots with significant differences in intensity/abundance between the sexes. Seven protein spots, which were prominent in 2‐DE patterns of male mice, but which showed very low intensities in females, were identified as selenium‐binding protein 2 (SBP2) also known as 56‐kDa acetaminophen‐binding protein. Edman degradation indicated that at least three of these protein spots represent N‐terminally truncated SBP2 variants. Furthermore, it was shown that the observed differences in SBP2 abundance correlate with sex differences in transcription of the gene encoding SBP2, selenbp2, as revealed by RT‐PCR and restriction digest as well as sequence analysis of the products. Since SBP2 has been described as the major target for acetaminophen in mouse liver cytosol, these findings are discussed with respect to their possible relevance for sex differences in acetaminophen‐mediated toxicity, which have been described in a variety of mammals including mice and rats.