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3 protocols using 5 sulphosalicylic acid

1

Plant Cell Culture Assay Reagents

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Murashige and Skoog medium, 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin, ethylenediaminetetraacetic acid (EDTA), triphenyl-tetrazolium chloride (TTC), 5-sulphosalicylic acid, 5,5-dithiobis(2-nitrobenzoic acid) (DTNB), deferoxamine, dimethyl sulfoxide (DMSO), acrolein, α-Tocopherol, Glutathione Reductase (GR), glutathione (GSH) and phenylmethylsulphonyl fluoride (PMSF) were obtained from Sigma-Aldrich. Ferrostatin-1, RSL3, Z-VAD-FMK and liproxstatin-1 were purchased from Selleckchem. 2’,7’-dichlorodihydrofluorescein diacetate (H2DCFDA) was purchased from Invitrogen. Ac-DEVD-AMC, Ac-DEVD-CHO and E64-d were from MedChemExpress. All other chemicals were from analytical or HPLC grade, and was purchased from Reanal, Hungary.
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2

Glutathione Reductase Activity Assay

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Glutathione reductase (GR), Ethylenediamine tetra acetic acid disodium salt, Nicotinamide adenine phosphate (β-NADPH), Pyrogallol, Trichloroacetic acid (TCA), 2-thiobarbituric acid (TBA), Sodium citrate, 1-chloro-2,4-dinitrobenzene, Reduced glutathione (GSH), Hydrogen peroxide (H2O2), 5,5-dithiobis-2-nitrobenzoic acid (DTNB), 5-sulphosalicylic acid, Hydrochloric acid, Nitric acid, trace metals grade perchloric acid, and n-butanol were procured from Sigma Aldrich, USA.
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3

Luminol-Based Chemiluminescence Assay

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Luminol (Sigma-Aldrich, MO, USA), CoCl2 (Univar, IL, USA), Na3PO4 7H2O (Mallinckrodt, Surrey, UK), NaOH (BDH, PA, USA), H2O2 (Chem-Supply Pty Ltd, South Australia, Australia), 5-sulphosalicylic acid (Sigma-Aldrich, MO, USA), ferrous ammonium sulphate (FeSO4(NH4)2SO4.6H2O) (England, UK), H2SO4 (Merck, VIC, Australia), xylenol orange (Sigma-Aldrich, MO, USA), Sorbitol (BDH, PA, USA), 0.45 µM PTFE captiva syringe filters (Agilent, CA, USA). Deionised water purified through a Milli-Q water purification system (Millipore, MA, USA) with a final resistance of 18.2 MΩ was used for all preparations unless mentioned otherwise.
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