Spectra max gemini xs fluorimeter

Testis tissue malondialdehyde (MDA) levels were measured using an MDA assay kit as per the manufacturer’s instructions (NWLSSTM Malondialdehyde Assay kit; Northwest Life Science Specialties LLC, Vancouver, WA, USA). Briefly, MDA reacts with thiobarbituric acid forming a colored complex which was measured at an absorbance of 532 nm by a Spectra Max 180 (Molecular Devices, Sunnyvale, CA, USA). The MDA level is expressed as μmole MDA per mg of wet tissue. Testis tissue ROS/RNS levels were detected using an OxiSelect in Vitro ROS/RNS Assay Kit per the manufacturer’s instructions (STA-347, Cell Biolabs, Inc., San Diego, CA, USA). Briefly, absorbance was measured at 480 nm excitation/530 nm emission with a SpectraMax Gemini XS Fluorimeter (Molecular Devices, Sunnyvale, CA, USA) as described previously [20 (link)]. The activities of superoxide dismutase (SOD) (706002, Cayman Chemical, Ann Arbor, MI, USA), glutathione peroxidase (GPx) (703102, Cayman Chemical, Ann Arbor, MI, USA), and catalase (707002, Cayman Chemical, Ann Arbor, MI, USA) in whole testis tissue supernatants were determined using commercially available kits per the manufacturer’s instructions. SOD, GPx, and catalase activities are expressed per milligram of protein.

Malondialdehyde (MDA) levels in testis tissues were assessed using reagents provided by Northwest Life Science Specialties LLC (Vancouver, WA, USA). Briefly, MDA reaction of a colored complex was measured at an absorbance of 532 nm using a kinetic spectrophotometer (Spectra Max 180, Molecular Devices, Sunnyvale, CA, USA). MDA levels are expressed as μmole of MDA/mg of wet testis tissue. Testicular ROS/RNS levels were quantified using an OxiSelect in Vitro ROS/RNS Assay Kit (STA-347, Cell Biolabs, Inc., San Diego, CA, USA) according to the manufacturer’s protocol. Fluorescence of dichlorofluorescein (DFC) was measured with a SpectraMax Gemini XS fluorimeter (Molecular Devices, Sunnyvale, CA, USA) at excitation/emission wavelengths of 480/530 nm as described previously [1 (link)]. Activities of testicular superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase were detected with commercially available kits (item no. 706002, 703102, and 707002, respectively, Cayman Chemical, Ann Arbor, MI, USA) as per the manufacturer’s instructions. Testicular SOD, GPx, and catalase values are expressed per milligram of protein.