Malondialdehyde mda assay kit

In summary, the hippocampal and cortical tissues were separated from the right hemi-brains (n = 6/group), homogenized in phosphate buffered saline (PBS) (mass ratio of 1:1) and were then divided into two parts, one for detecting SOD/MDA, and the other for detecting protein by western blot analysis. PBS (mass ratio of 1:10) was added to the homogenates and homogenization was repeated using a Polytron homogenizer. The homogenates were centrifuged at 3,000 r/min for 10 min at 4°C and the supernatants were collected. The SOD activity and MDA levels were determined according to the manufacturers’ instructions for Total Superoxide Dismutase (T-SOD) assay kit (Hydroxylamine method) or Malondialdehyde (MDA) assay kit (TBA method) from Nanjing Jiancheng Bioengineering Institute.

The activity of SOD and the contents of MDA and ROS in the perihematoma tissues of rat brain in each group were determined by Superoxide Dismutase (SOD) assay kit, Malondialdehyde (MDA) assay kit, and Reactive oxygen species (ROS) assay kit (Nanjing Jiancheng Bioengineering Institute, China) according to the operating instructions.

Juglone (purity≥98%) was obtained from Shanghai Yuanye Biotechnology Co., LTD (Shanghai, China). Cis-platinum was purchased from Qilu Pharmaceutical Co., LTD (Jinan, China). The primary antibodies for PI3K, p-PI3K, Akt, p-Akt, caspase-3, cleaved caspase-3, Bcl-2, Bax, Cyt c, and β-actin were obtained from Santa Cruz Biotechnology Inc. (Dallas, CA, USA). 1,3-Dicaffeoylquinin acid (1,3-diCQA, an activator of PI3K/Akt), N-acetyl-L-cysteine (NAC, a ROS inhibitor), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), trypsin, DAPI reagents, and sodium dodecyl sulfate (SDS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Matrigel was obtained from BD Biosciences (San Jose, CA, USA). High glucose DMEM medium and fetal bovine surum (FBS) were provided by Gibco (Grand Island, NY, USA). ROS detection kit was obtained from KeyGen BioTECH (Nanjing, China). Malondialdehyde (MDA) assay kit, glutathione peroxidase 4 (GPX4) assay kit and superoxide dismutase (SOD) assay kit were purchased from Jiancheng Bioengineering Institute (Nanjing, China).

Antimycotics (#15240062), fetal bovine serum (FBS, #10091148), RPMI 1640 medium (#A1049101), and Dulbecco's modified Eagle's high glucose medium (#11965118) were purchased from HyClone (HyClone, Logan, UT, USA). Cell Counting Kit-8 (CCK-8, #CK04) was obtained from Dojindo (Dojindo Laboratories, Kumamoto, Japan). The malondialdehyde (MDA) assay kit (#A003-1-2) and reduced glutathione (GSH) assay kit (#A006-2-1) were purchased from Jiancheng (Jiancheng, Nanjing, China). The DAPI staining solution kit (#MA0127) and zinc protoporphyrin-9 (#MB4231) were purchased from Meilui (Meilui, Dalian, China). Liperfluo (#L248) was purchased from Dojindo (Dojindo Laboratories, Shanghai, China). 2′,7′-Dichlorodihydrofluorescein diacetate (DCFH-DA, #S0033S), Annexin V (#C1062M-1), and Propidium Iodide (PI, #C1062M-3) were purchased from Beyotime (Beyotime, Shanghai, China). For western blot analysis, antibodies against HO-1 (#ab13243), GPX4 (#ab40993), Nrf2 (#ab137550), and β-actin (#ab8226) were purchased from Abcam (Abcam, Cambridge, MA). The Iron Assay Kit (#MAK025), dimethyl sulfoxide (DMSO, #D2650), curcumin (#C1386), z-VAD-fmk (#V116), ferrostatin-1 (#SML0583), and deferoxamine (#D9533) were purchased from Sigma (Sigma, St Louis, USA). These compounds were dissolved in DMSO (at final concentrations less than 0.1% (v/v)).