Adipose tissue serves as a major endocrine organ with a profound influence on metabolism by secreting and regulating numerous molecules, hormones and adipokines. Many proteins activate intracellular pathways that promote the development of insulin resistance (IR); however, the role of specific proteins in adipose tissue dysfunction is not well defined. The objective was to identify proteins in adipose that are linked to IR and to cows' metabolic status. Adipose tissue biopsies were obtained from 8 multiparous cows at −17 and +4 d relative to parturition. Proteins were analyzed by intensity based, label-free quantitative shotgun proteomics at Weizmann Institute of Science (Rehovot, Israel). Proteins were extracted and subjected to in-solution tryptic digestion. This was followed by nanoflow liquid chromatography coupled to high-resolution tandem mass spectrometry (nanoLC-MS/MS). Quantitative data were extracted using the Genedata Expressionist data analysis package and proteins identified using the Mascot search engine. Cows were previously divided to those with IR or insulin-sensitive (IS) adipose based on phosphorylation of protein kinase B (Akt) in response to insulin stimulation. Proteomics data, after logarithmic transformation, were analyzed by 2-way ANOVA to measure the effects of time (prepartum vs. postpartum), subgroup (IR vs. IS) and their interaction. Body weight (BW) differences were analyzed with GLM of SAS. It was found that cows with IR adipose lost more BW postpartum compared with IS cows. Proteomic analysis revealed 586 proteins in adipose tissues. Comparing IR to IS adipose showed that 18.9% of proteins were differentially expressed (fold change (FC) > 1.5 and P < 0.05). The expression of 106 proteins were increased, whereas only 5 were decreased, in IR adipose compared with IS. The abundance of several proteins related to lipolysis was increased in IR adipose compared with IS: hormone-sensitive lipase (FC = 6.8, P < 0.03), perilipin (FC = 1.5, P < 0.05), and monoglycerol-lipase (FC = 8.2, P < 0.0003). This is in accordance with the elevated lipolysis in IR adipose. These proteins could be used as novel biomarkers to identify IR cows, which may indicate of the metabolic status of the peripartum dairy cow.