We performed label-free micro-data-independent acquisition (µDIA) to assay brain proteins from a severe model of Alexander disease (AxD, lethal at postnatal day ~35). Of the 5,005 proteins quantified, we observed upregulation of adipocytokine signaling, PPAR, insulin resistance, and glutathione pathways with concomitant downregulation of platelet activation, ether lipid synthesis, and steroid biosynthesis pathways. To validate these DIA results, we setup a targeted parallel reaction monitoring-mass spectrometry assay (PRM-MS) for selected proteins with altered relative concentrations. The PRM-MS assays confirmed the upward or downward relative concentration differences in 16/16 (100%) of the proteins targeted, with 15/16 (94%) of these proteins reaching statistical significance (P-value < 0.05).