Oxiselect tbars assay kit

Thiobarbituric Acid Reactive Substances (TBARS) has been used for MDA assay for screening and monitoring lipid peroxidation. MDA quantification was performed using OxiSelectTM TBARS Assay Kit (STA-310, Cell Biolabs, Inc.) according to manufacturer’s instructions.

Lipid peroxidation was quantified with OxiSelect^TM TBARS Assay Kit (STA-330, Cell Biolabs, Inc., San Diego, CA, USA) by following the protocol set by the manufacturer. Samples were allowed to react with thiobarbituric acid (TBA), and the resultant mixture was measured at 532 nm.

Malondialdehyde (MDA) of samples was determined with the OxiSelectTM TBARS Assay Kit (Cell Biolabs, Inc., San Diego, CA, USA). A 100 μL sample was mixed with 100 μL 10% sodium dodecyl sulphate (SDS) lysis solution in a microcentrifuge tube and incubated for 5 min at room temperature. Then, 1X 2-thiobarbituric acid (TBA) diluent (250 μL) was added. The mixture was allowed to incubate at 100 °C for 50 min. After that, the tube was removed and placed into an ice bath for 5 min. The tube was centrifuged at 9000 rpm, 4 °C for 15 min. The supernatant of sample (200 μL) was transferred to a 96-well microplate. Absorbance was measured at 532 nm. MDA standard solution was used as standard. TBARS of liver supernatant were expressed as nanomole of MDA equivalents per gram protein.

Concentrations of cytokines and oxidative modification products were determined in 10% (weight/volume) lung homogenate prepared using 0.1 M ice-cold phosphate buffer (PBS, pH 7.4) by Polytron homogenizer PT 1200 E (5-times for 25 s, 1200 rpm; Kinematica AG, Switzerland). Homogenates were then frozen 3 times and centrifuged (12,000 rpm, 15 min, 4 °C). Final supernatants were then stored at −70 °C until the analysis was performed.
Concentrations of IL-6, IL-1β, IL-10 (USCN Life Science Inc., Wuhan, China), and RAGE (MyBioSource, San Diego, CA, USA) were quantified using rabbit-specific ELISA kits according to the manufacturers’ instructions. Data were expressed in pg/mL.
Protein oxidative damage was determined using OxiSelect^TM Nitrotyrosine ELISA Kit (Cell Biolabs, Inc., San Diego, CA, USA). Data were expressed in 3-nitrotyrosine nanomole concentration (nM, 3NT). Lipid oxidative damage expressed by the concentration of thiobarbituric acid reacting substances (TBARS) was determined by OxiSelect^TM TBARS Assay Kit (Cell Biolabs, Inc., San Diego, CA, USA). Data were expressed as malondialdehyde in micromole concentration (μM MDA).
Total antioxidant capacity (TAC) was determined using an ELISA kit according to the manufacturer’s instructions (Cell Biolabs, Inc., San Diego, CA, USA). Data were expressed as micromole concentration of copper reducing equivalents (μM CRE).

Adipose tissue lipid peroxidation was measured with OxiSelect^TM TBARS Assay Kit (#STA-330, Cell Biolabs, Inc. San Diego, CA, USA), as per the guidelines set by the manufacturer. Briefly, samples were incubated with lysis buffer and allowed to react with thiobarbituric acid (TBA). Cooled samples were centrifuged to collect the supernatant, and absorbance was measured at 532 nm.

Lipid peroxidation levels in plasma were estimated by measuring thiobarbituric acid reactive substances (TBARS) and expressed in terms of malondialdehyde (MDA) content, which is the product of lipid peroxidation by using the OxiSelect^TM TBARS assay kit (Cell Biolabs Inc., San Diego, CA, USA).