Mda assay kit tba method

MDA content and SOD activity were measured in renal tissues and HK2 cells to assess the level of oxidative stress. The intracellular concentration of MDA was detected using a MDA assay kit (TBA Method, Nanjing Jiancheng Bioengineering Institute) and was reported as nmol/mg of extracted protein. SOD activity was detected using the Total Superoxide Dismutase (T-SOD) assay kit (Hydroxylamine method, Nanjing Jiancheng Bioengineering Institute) and was reported as U/mg protein.

To assess the oxidative stress in intracellular and periodontal tissues, the levels of ROS, malondialdehyde (MDA), and superoxide dismutase (SOD) were detected.
The total intracellular ROS levels were determined with the ROS Assay Kit (Beyotime, China). Briefly, cells were incubated with DCFH-DA reagent for 20 min at 37°C after treatment. The intracellular fluorescence intensity was then examined under a fluorescent inverted microscope (Olympus, Japan).
Mitochondrial-derived ROS levels in periodontal tissue were measured using the Mito SOX Red kit (Thermo Fisher Scientific, United States). The Mito SOX red reagent can be oxidized by superoxide to show red fluorescence and thus assess the ROS content. The staining process was carried out as directed. The Mito SOX red fluorescence of the sections was detected by confocal microscopy.
The SOD typed assay kit (Hydroxylamine method) and MDA assay kit (TBA method) (Nanjing Jiancheng Bioengineering Institute, China) were used to detect intracellular and serum levels of MDA and SOD activity. The experimental procedures were carried out strictly according to the instructions.

MDA content was analyzed as described by previous report [29 (link)]. MDA assay kit (TBA method) (A003–1, Nanjing Jiancheng Bioengineering Institute (Nanjing, China)) was used according manufacturer’s instructions. MDA reacted with thiobarbituric acid (TBA) formed a red-complex compound featuring absorbance at 532 nm. MDA results were expressed in nmol/mL in serum and tissues.

The cell culture supernatants were collected for measuring superoxide dismutase (SOD) and malondialdehyde (MDA) after OGD. The cell culture supernatants were centrifuged at 3000 rpm for 10 min at 4 °C and then collected for determination. Thereafter, the SOD levels were measured using the total SOD determination kit (WST-1 method) (Jiancheng Biotechnology, Nanjing, China). The MDA levels were tested using an MDA assay kit (TBA method) (Jiancheng Biotechnology, Nanjing, China) according to the manufacturer’s instructions.

ORO was purchased from Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). Superoxide Dismutase (SOD) assay kit (WST-1 method), catalase (CAT) assay kit, MDA assay kit (TBA method), and Triglyceride (TG) assay kit were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). BCA Protein Assay Kit was purchased from Beyotime biotechnology (Shanghai, China). α-linolenic acid was purchased from sigma Aldrich Trading Co., Ltd. (Shanghai, China). 5-Hydroxy-1,4-naphthalenedione and 2, 7-dichlorofluorescein diacetate were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). MiniBEST Universal RNA Extraction Kit, PrimeScript^TM RT Master Mix (Perfect Real Time), and TB Green^® Premix Ex Taq^TM II (Tli RNaseH Plus) were purchased from Takara (Dalian, China). All other chemicals were of analytical grade and used without further purification.