Malonaldehyde bis dimethyl acetal

Bovine serum albumin (BSA), Giemsa, 2,2-Diphenyl-1-picrylhydrazyl (DPPH), sodium nitroprusside, quercetin, sulphanilamide, phosphoric acid, tri sodium citrate, ammonium molybdate, sodium chloride, hydrogen peroxide_, thiobarbituric acid (TBA), trichloroacetic acid (TCA), hydrochloric acid, 2,4-Dinitrophenylhydrazine (DNPH), malonaldehyde-bis-dimethyl acetal (MDA) and May-Grunwald’s stain were procured from Sigma-Aldrich ( St Louis, USA). Rat COX-2 ELISA kit was obtained from Genxbio health sciences Pvt Ltd, India. Rat IL-6 ELISA kit and Rat IL-10 ELISA kit was procured from Ray Biotech Pvt Ltd, India. All other chemicals and reagents used were of analytical grade.

δ-aminolevulinic acid (δ-ALA), DL-dithiothreitol (DTT), rabbit metallothionein-I, o-phenylendiamine, thiobarbituric acid (TBA) and malonaldehyde bis- (dimethyl acetal) were obtained from Sigma (St. Louis, MO, USA); mouse anti-metallothionein-I/II, and peroxidase-conjugated to goat anti-mouse IgG were purchased from Dako Corporation (Carpinteria, CA, USA).

24 hours following cocaine-induced CPP, animals were euthanized and their brains were immediately removed. PFC and NAc were microdissected, homogenized and analyzed for OS. The extent of lipid peroxidation in animal brain tissue was estimated colorimetrically using the thiobarbituric acid reactive substances (TBARS) method as we have previously described^{16 (link)}. Briefly, 0.1 ml aliquots of tissue homogenates were treated with 2 ml of (thiobarbituric acid (TBA)–TCA–HCl 1:1:1 ratio) reagent containing: thiobarbituric acid 0.37% (Sigma, St. Louis, USA) - TCA 15% (Baker, NJ, USA) - HCI 0.25 N (1:1:1 ratio). Samples were heated to 95 °C for 30 min, cooled on ice and centrifuged (12,000 rpm, 10 min). The clear supernatant fluids were measured at 535 nm with a microplate reader (Bio-TEK ELx, Winooski, VT, USA). A malonaldehyde bis (dimethyl acetal) (Sigma, St. Louis, USA) standard curve was prepared under the same experimental conditions. Calculations were made on the basis of standard curves and presented as µmol malondialdehyde (MDA)/g protein.

WPI (>91.5%) was purchased from Hilmar Industries (Hilmar, CA, USA). TNTs (purity > 99.9%, OD ≈ 20 nm) and TNPs were obtained from the Beijing Dk Nano S&T, Ltd. (Beijing, China). 2-Thiobarbituric acid (TBA), trichloroacetic acid (TCA), and malonaldehyde bis (dimethyl acetal) were purchased from Sigma-Aldrich (St Louis, MO, USA). 2,2-Diphenyl-1-picrylhydrazyl (DPPH) was purchased from Aladdin Reagent Co. (Shanghai, China). Pathogenic bacteria were obtained from BNCC Biological Technology Co., Ltd. (Nanjing, China). Ultrapure water was prepared from a Milli-Q water purification system (Millipore, Billerica, MA, USA) and was used throughout the experiments. Other reagents obtained in this study were of analytical grade.

Tert-butyl hydroperoxide (t-BHP), sodium dodecyl sulfate (SDS), phosphotungstic acid, buthylated hydroxytoluene, 2-thiobarbituric acid and malonaldehyde bis(dimethyl acetal), the Bicinchoninic protein determination assay and the anti-α-tubulin were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). The following were purchased from Abcam Inc., (Cambridge, MA, USA): rabbit polyclonal anti-PRDX1, mouse monoclonal anti-PRDX6, mouse monoclonal anti-4-Hydroxynonenal (4-HNE), rabbit polyclonal anti-catalase, and 8-hydroxy-deoxyguanosine (8-OHdG). Anti-thioredoxin 1 antibody was purchased from Cell Singaling Thecnology (Danvers, MA, USA), Polyvinylidene fluoride (PVDF) membranes (0.22 µm pore size; Osmonics Inc., Minnetonka, MN, USA), donkey anti-rabbit IgG and goat anti-mouse IgG, both conjugated to horseradish peroxidase (Cedarlane Laboratories Ltd., Hornby, ON, Canada), an enhanced chemiluminescence kit (Lumi-Light; Roche Molecular Biochemicals, Laval, QC, Canada) and radiographic films (Denville Scientific, Inc., Saint-Laurent, QC, Canada) were also used for immunodetection of blotted proteins. Other chemicals were used at the reagent level.