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Significantly, all synthesized compounds have been tested for their promising antioxidant activities via utilization of 1,1-biphenyl-2-picrylhydrazyl as a free radical scavenging reagent.
To further prove that respiration and accompanying ROS production is a major cause of apoptosis during colony development we used reduced glutathione (GSH) as a scavenging reagent: Colonies of wild-type and oxa1 deletion strains were grown on SCGlu agar plates with or without GSH and cell survival was determined after 36 hours in a clonogenic assay.
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To determine GSSG, a duplicate of each sample was previously incubated with 3 mmol/l of 1-methyl-2-vinylpyridinium triflate, a thiol-scavenging reagent, which rapidly scavenges reduced form of GSH without interfering with the enzyme activity.
Tissue was homogenized by sonication with or without the thiol-scavenging reagent 1-methyl-2-vinylpyridinium trifluoromethanesulfonate (M2VP).
Samples for GSSG measurement were immediately mixed with thiol-scavenging reagent M2VP (1-methyl-2-vinyl-pyridium trifluoromethane sulfonate) after separation.
Scavenging mechanisms including various antioxidant reagents and enzymes have been found in cells to eliminate free radicals.
Hydrogen peroxide scavenging capacity of SSME was studied by FOX reagent method in which oxidation product of Fe2+ binds to xylenol orange.
The scavenging ability on DPPH radicals was calculated as follows: Scavenging ability on DPPH radicals = [(A1 - A2)/A1] × 100, where A1 is the absorbance of the control (containing all reagents except the sample extract), and A2 is the absorbance of the sample extract.
All determination was performed in triplicate and results were performed in triplicate and results are reported as scavenging effect versus concentration in Figure 2. TPC were determined using the Folin-Ciocalteu reagent method described by Singleton and Rossi [ 12].
The scavenging ability of hydroxyl radicals was calculated using the following equation: Scavenging ability on hydroxyl radicals = [(A1 - A2)/A1] × 100, where A1 is the absorbance of the control reaction (containing all reagents except the sample extract), and A2 is the absorbance of the sample extract.
Iron is a key limiting reagent during in vivo bacterial infection, and multiple systems have been identified in SA for binding and scavenging iron from mammalian host iron-binding proteins (IBPs) such as lactoferrin, transferrin and hemoglobin [40] [42].
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com