The periparturient period is characterized by increased oxidative stress status in dairy cows. In non-ruminants, a chronic state of excessive fat deposition is accompanied by inflammation and oxidative stress. The objective was to investigate if prepartal body condition score (BCS) is associated with plasma and adipose tissue biomarkers of oxidative stress in Holstein cows. Twenty 2 multiparous Holstein cows were divided by BCS before parturition (−30 d prepartum) into a BCS ≤ 3.25 (LoBCS, n = 11) or BCS ≥ 3.50 (HiBCS, n = 11) group. Blood sampled from the coccygeal vessel at −30, −15, 7, 10, and 30 d relative to calving date was used for oxidative stress and inflammation biomarkers analysis. Adipose tissue obtained from each group on d −15, 7 and 30 relative to calving date was used for RT-PCR analysis. The statistical model in SAS included the fixed effect of treatment, time and its interaction. There was no difference in prepartal DMI and milk yield between groups (P > 0.05). A treatment × time interaction (P < 0.05) was observed for postpartal DMI due to greater responses in LoBCS cows. Although there was no overall treatment or interaction effect (P > 0.05) for plasma myeloperoxidase and haptoglobin, the fact that ferric-reducing ability (antioxidant capacity) increased between −10 and 7 d (interaction P = 0.01) and reactive oxygen metabolites between −10 and 15 to 30 d (interaction P < 0.01) in HiBCS cows suggested a chronic inflammatory and oxidative stress state. The greater (P < 0.05) overall concentration of carotene in LoBCS cows suggested they had better antioxidant status. Cows with HiBCS had greater abundance of cullin 3 (CUL3) (P < 0.05) and lower overall abundance of factor erythroid 2-like 2 (NFE2L2). However, HiBCS cows had greater (P < 0.05) abundance of genes associated with glutathione metabolism including glutathione peroxidase 1 (GXP1) and glutathione reductase (GSR). Regardless of BCS, mRNA abundance of NEF2L2 decreased and KEAP1 increased from 7 to 30 d after parturition. Overall, the data suggest a more pronounced systemic and localized oxidative stress status in cows with prepartal BCS ≥ 3.50.