The aim of this study was to evaluate the influence of applying a 4-week instead of an 8-week dry period to dairy cows on the proteome of colostrum (first sample) and of transition milk (the fifth postpartum milk sample). Individual milk serum samples of colostrum and transition milk were analysed from 12 Swedish Holstein (SH) and 12 Swedish Red (SR) cows. Samples were prepared by filter-aided sample preparation and dimethyl labelling and analysed by liquid chromatography tandem mass spectrometry. Shortening the dry period resulted in upregulation of 18 proteins in colostrum and transition milk of SR, whereas no statistical differences were found for SH colostrum and transition milk. These upregulated proteins may reflect a specific immune response in the SR samples that was reflected in colostrum as well as in transition milk. Upregulated proteins in colostrum seemed to reflect increased mammary epithelial cell proliferation in the periparturient period when a 4-week dry period was applied. The proteome data indicate that a dry period of 4 weeks to SR cows may not be sufficient for complete regeneration of the mammary epithelium.