During periparturient metabolic stress, excessive adipose lipolysis is a strong predisposing factor for postpartum complications. Linoleic acid is the most abundant polyunsaturated fatty acid (FA) in adipose tissue (AT) of dairy cows and is preferentially released during lipolysis. In the adipocyte, oxidized linoleic acid metabolites (OXLAM) are products of enzymatic and non-enzymatic pathways. Among OXLAM, 13-hydroxyoctadecadienoic acid (HODE) was identified as a lipolysis product, but its role in modulating lipid mobilization in AT is unclear. We hypothesize that 13-HODE reduces lipolytic response to adrenergic stimulation and enhances FA uptake in adipocytes. Subcutaneous AT explants collected from dairy cows (n = 7) at 10 ± 2 d postpartum and cultured bovine adipocytes (n = 4) were incubated with physiological concentrations of 13-HODE: 0 nM (CO), 50 nM (HL), or 200 nM (HH) for 4 h. To stimulate lipolysis, explants were then exposed to the β adrenergic agonist isoproterenol (BAS 0M; ISO, 10⁻⁶ M) for 3 h. Lipolytic responses were evaluated by measuring glycerol release. To evaluate the effect of 13-HODE on lipogenic capacity, adipocytes were treated with insulin (5 μg/mL, 1 h) and the rate of FA uptake was measured with the QBT FA uptake assay. The statistical model included the random effect of animal and the fixed effect of treatment, insulin or ISO stimulation and their interaction. Exposure of AT explants to HL and HH for 4 h did not induce a lipolytic response compared with CO (P = 0.62). Glycerol release upon adrenergic stimulation was reduced in HL+ISO and LL+ISO compared with CO+ISO (P < 0.05). Incubation with 13-HODE did not affect FA uptake in cultured adipocytes (P = 0.43). β-adrenergic agonist stimulation enhanced gene expression of 13-HODE targets GPR132 and testicular orphan nuclear receptor 4 (TR4), key regulators of lipid trafficking in adipocytes. These data suggest that 13-HODE is a modulator of lipolysis but not lipogenesis in AT. Future studies will evaluate the mechanisms by which 13-HODE signals through GPR132 and TR4 to exert its effects on AT lipolytic responses.