The content of reactive oxygen species (ROS) was determined using the Oxiselect™ assay kit (Cell Biolabs Inc., San Diego, CA).
Oxiselect assay kit
Manufactured by Cell Biolabs
Sourced in United States
The Oxiselect™ assay kit is a quantitative tool that measures oxidative stress levels in biological samples. It provides a sensitive and reliable method to detect and quantify various reactive oxygen species (ROS) and oxidative damage markers.
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Lab products found in correlation
8 protocols using oxiselect assay kit
1
ROS Quantification via Oxiselect Assay
2
Quantifying Total Antioxidant Capacity
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The total antioxidant capacity was assessed using the Oxiselect™ assay kit (Cell Biolabs, San Diego, CA). The copper ion reagent (Cu2+) or a known concentration of uric acid was added to the samples. Upon reduction, Cu2+ reacts with a chromogenic reagent that produces an orange colored product. The absorbance of this product at 490 nm was measured using a MultiSkan FC spectrometer (Thermo Fisher Scientific, Waltham, MA). The results were expressed as mM of uric acid/mg protein.
3
Measuring Hepatic Oxidative Stress
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Hepatic ROS level was measured by using OxiSelect Assay Kit (STA-347, Cell Biolabs, Inc., San Diego, CA, USA). Liver tissue was cut on dry ice, weighed, resuspended in PBS at 50 mg/mL, and homogenized. Samples were centrifuged at room temperature for 5 min at 10,000× g, supernatants transferred to a new tube, and all samples were assayed in duplicate, following manufacturer’s instructions.
4
Measuring Anthocyanins and Antioxidants in Wheat
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The total anthocyanins were extracted and measured by a spectrophotometric method (Abdel-Aal & Hucl, 1999) using a V650 spectrophotometer (Jasco, Japan). The results are expressed as mg/kg DM on the basis of the cyanidin 3-glucoside standard calibration curve. The total polyphenolic content (TPC) of bread wheat flour, GPP and cakes was assessed on 80% methanol extracts by the Folin-Ciocalteu method as described in Nakov et al. (2018a) (link) and was expressed as mg gallic acid equivalent (GAE)/g dry matter (DM). The antioxidant capacity was determined with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, according to Nakov et al. (2018a) (link), and by the ferric reducing antioxidant power (FRAP) with the OxiSelect™ Assay Kit (Cell Biolabs, Inc., San Diego, CA, USA). The DPPH and the FRAP results are presented as μmol Trolox equivalents (TE)/g DM.
5
Measuring Anthocyanins and Antioxidants in Wheat
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The total anthocyanins were extracted and measured by a spectrophotometric method (Abdel-Aal & Hucl, 1999) using a V650 spectrophotometer (Jasco, Japan). The results are expressed as mg/kg DM on the basis of the cyanidin 3-glucoside standard calibration curve. The total polyphenolic content (TPC) of bread wheat flour, GPP and cakes was assessed on 80% methanol extracts by the Folin-Ciocalteu method as described in Nakov et al. (2018a) (link) and was expressed as mg gallic acid equivalent (GAE)/g dry matter (DM). The antioxidant capacity was determined with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, according to Nakov et al. (2018a) (link), and by the ferric reducing antioxidant power (FRAP) with the OxiSelect™ Assay Kit (Cell Biolabs, Inc., San Diego, CA, USA). The DPPH and the FRAP results are presented as μmol Trolox equivalents (TE)/g DM.
6
Testicular Antioxidant Enzyme Evaluation
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Testicular level of superoxide dismutase (SOD) was estimated using rat ELISA assay kit (Cusabio Biotech Company, Wuhan, China). Testicular levels of catalase (CAT), glutathione peroxidase (GPx) and nitric oxide (NO) were measured with rat ELISA assay kits ((MyBioSource, San Diego, CA, United States) following the manufacturer’s protocol. Testicular levels of malondialdehyde (MDA), the lipid peroxidation marker, were measured by commercially available rat ELISA kits (Elabscience, Wuhan, China) according to the manufacturer’s instructions. Testicular and seminal total antioxidant capacity (TAC) were assessed using Cell Biolabs OxiSelect™ assay kits (San Diego, Inc., CA, USA), according to the manufacturer’s instructions.
7
Antioxidant Enzyme and Lipid Peroxidation Analysis in Testicular Tissue
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Testicular homogenates were used to evaluate antioxidant enzyme activities, nitric oxide (NO) concentrations, and lipid peroxidation levels (malondialdehyde; MDA activity). Superoxide dismutase (SOD) was measured using commercial ELISA kits (Cusabio Biotech Co., Ltd., Wuhan, China), according to the manufacturer's protocol. Catalase (CAT), glutathione peroxidase (GPx), and total NO levels were estimated using rat ELISA kits (MyBioSource, San Diego, CA, USA), according to the manufacturer's instructions. The levels of lipid peroxidation were determined using MDA assay kits (Elabscience Biotechnology Co., Ltd., Wuhan, China). Total antioxidant capacity (TAC) in semen and testicular homogenate was estimated using Cell Biolabs OxiSelect assay kits (San Diego, Inc., CA, USA), according to the manufacturer's instructions.
8
Antioxidant Capacity Evaluation Protocol
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The total antioxidant capacity (TAC), superoxide dismutase (SOD), and catalase activity were measured using OxiSelect™ assay kits (Cell Biolabs Inc., San Diego, CA, USA) according to the manufacturer’s protocol. Non-conditioned medium as control, ASC-CM and AMSC-CM were assessed for TAC, SOD, and CAT activity levels. Culture medium of the control, ASC-CM, and AMSC-CM groups before and after IVC were assessed for SOD and CAT activity levels. The results of each colorimetric assay were assessed using measured absorbances at 490 nm for TAC and SOD activity, and 520 nm for CAT activity.
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