Gsh elisa kit

All kits used for this part were rat-specific. MDA ELISA kit for rats was used to measure levels of malondialdehyde (MDA) in the liver homogenates (#MBS268427, MyBioSource, CA, USA). GSH ELISA kit (#MBS265966; MyBioSource, CA, USA), SOD ELISA kit (#MBS036924, MyBioSource, CA, USA), CAT ELISA kit (#MBS726781, MyBioSource, CA, USA), IL-6 ELISA kit (#MBS269892, MyBioSource, CA, USA), HO-1 ELISA kit (MBS764989, MyBioSource, CA, USA) and TNF-αELISA kits (#MBS2507393, MyBioSource, CA, USA) were used to measure the levels of glutathione (GSH), superoxide dismutase (SOD), catalase (CAT), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the liver homogenates. G6PC ELISA kit (#MBS097902 MyBioSource, CA, USA), FBP1 ELISA kit (#MBS931493, MyBioSource, CA, USA), and GLY ELISA kit (#MBS1600418; MyBioSource, CA, USA) were used to measure the levels of glucose-6 phosphatase (G-6-Pase), fructose-1,6-bisphosphatase 1 (PBP1), and glycogen in the liver homogenates. The TransAM Nrf2, NF-E2-related factor 2 (Nrf2 DNA binding ELISA) kit (#50296, ActiveMotif, Tokyo, Japan) and TransAM NF-κB p65 ELISA kits (#40096, ActiveMotif, Tokyo, Japan) were used to measure the total and cytoplasmic levels of Nrf2 and NF-κB p65 in the homogenates. All procedures were conducted according to each manufacturer’s instructions for 8 samples/groups.

Gestational, 1 h post-calving, and lactational cow serum were analyzed by Novus International, Inc. for thiobarbituric acid reactive substrates (TBARS) concentration using a commercially-available kit (Cayman Chemical, Ann Arbor, MI), as well as glutathione peroxidase (GPx), reduced glutathione (GSH), and oxidized glutathione (GSSG) concentration using an ELISA assay (GSH-Px ELISA kit, GSH ELISA kit, and GSSG ELISA kit, MyBioSource, Inc., San Diego, CA). Samples were analyzed in duplicate using a microplate reader (Epoch 2, BioTek, Winooski, VA) at 532 nm for TBARS assay and 450 nm for GPx, GSH, and GSSG assays. Intraassay and interassay CV for TBARS, GPx, GSH, and GSSG were < 3.0% and < 2.7%, respectively. Serum protein concentrations were used to report GPx, GSH, and GSSG concentrations relative to serum protein to ensure these oxidative stress marker concentrations were not confounded by serum dilution. Serum was analyzed in duplicate for protein concentration using a commercially available Coomassie (Bradford) Protein Assay Kit (Thermo Scientific, Waltham, MA). Intraassay and interassay CV were 5.2% and 2.9%, respectively.

The lysate from DRG was prepared by incubating the DRGs with 1% of protease inhibitor cocktail III (Fisher Scientific, St. Louis, MI, USA) for 1 h. The protein contents in the supernatant were quantified using a Bradford protein assay.
The oxidative stress parameters were measured using mice malondialdehyde (MDA) ELISA kit (MyBioSource MBS263626, San Diego, CA, USA), a mice glutathione (GSH) ELISA kit (MyBioSource MBS267424, San Diego, CA, USA), and a mice superoxide dismutase enzymes (SOD) ELISA kit (MyBioSource MBS034842, San Diego, CA, USA), following the manufacturer’s instructions.
The pro-inflammatory biomarkers were measured using a mouse nuclear factor KB ELISA Kit (MyBioSource MBS043224, San Diego, CA, USA) and the tumor necrosis factor kit assay (MyBioSource MBS825075, San Diego, CA, USA), following the manufacturer’s instructions.