Abts free radical scavenging capacity assay kit

The in vitro concentration-dependent antioxidant activity of allantoin was evaluated using the DPPH Free Radical-Scavenging Capacity Assay kit (Solarbio Technology Co., Ltd., Beijing, China), the ABTS Free Radical-Scavenging Capacity Assay kit (Solarbio Technology Co., Ltd.), and the Total Antioxidant Capacity (T-AOC) Assay kit (Solarbio Technology Co., Ltd.). Allantoin was used at doses of 0.01, 0.025, 0.05, 0.1, and 0.15 mg/mL in the assays. An authentic antioxidant (vitamin C) used at the same doses as allantoin served as the control to confirm the utility of the assays. All measurements were completed using three independent biological replicates.

Choline chloride (ChCl, 98%, CID: 6209), urea (99%, CID: 1176), citric acid (≥99.5%, CID: 311), malic acid (99% CID: 525), ethylene glycol (98%, CID: 174), malonate (99.5%, CID: 9084), glycolic acid (98%, CID: 757), and ethanol (EtOH, 99.9%, CID: 702) were of analytical grade and acquired from Macklin Biochemical Co., Ltd. (Shanghai, China). 1,2-propanediol (AR, ≥99.5%, CID: 1030), vitamin C (VC), DPPH free radical scavenging capacity assay kit, and ABTS free radical scavenging capacity assay kit were purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China). Methanol (CID: 887), acetic acid (CID: 176), water (CID: 962), vitexin (CID: 5280441), and isovitexin (CID: 162350) were of chromatographic grade and acquired from Aladdin (Shanghai, China).

Polyethyleneimine (PEI, M.W. 10,000, 99%), manganese chloride tetrahydrate (MnCl₂·4H₂O), and 3,3′,5,5’-Tetramethylbenzidine (TMB) were purchased from Aladdin Reagent Co., Ltd. (Shanghai, China). Hydrogen peroxide (H₂O₂, 30%), hydrochloric acid (HCl), sodium acetate (NaAc), acetic acid (HAc), and ethanol (C₂H₅OH) were obtained from Sinopharm Chemical Reagent Co., Ltd. (Shanghai, China). Titanium (IV) sulfate (Ti(SO₄)₂) was provided by Macklin (Shanghai, China). All the chemicals were of analytical grade and used directly without further purification.
Curcumin was purchased from MedChemExpress (Monmouth Junction, NJ, USA). Lipopolysaccharide (LPS) was provided by Sigma Aldrich (St. Louis, MO, USA). TNF-α (#ab1793), iNOS (#ab178945) and IL-10 (#ab9969) were obtained from Abcam (Cambridge, MA, USA). H2AX (#10856-1-AP) and β-actin (#66009-1-Ig) were obtained from Proteintech (Wuhan, China). Calcein AM, Propidium Iodide, the Total Superoxide Dismutase Assay Kit with WST-8, the Reactive Oxygen Species Assay Kit, and the Lipid Peroxidation MDA Assay Kit were obtained from Beyotime (Shanghai, China). The ABTS Free Radical Scavenging Capacity Assay Kit, DPPH Free Radical Scavenging Capacity Assay Kit, and CCK-8 Cell Proliferation and Cytotoxicity Assay Kit were purchased from Solarbio (Beijing, China).