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2 protocols using β d glucose

1

Antioxidant and Cytotoxicity Assays

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Standards of ascorbic acid, 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), potassium persulphate, 6-hydroxy-2,5,7,8-tetramethyl-chroman-2-carboxylic acid (Trolox), and 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), thiazolyl blue tetrazolium blue (MTT), bromophenol blue, and Tris–HCl were purchased from Sigma–Aldrich Chemical Co. (St. Louis, MO, USA). β-d-(+)-Glucose was purchased from ChromaDex (Irvine, CA, USA). Dulbecco’s Modified Eagle Medium (DMEM, high glucose + GlutaMAX), fetal bovine serum (FBS), Dulbecco’s phosphate-buffered saline (DPBS) were provided by Gibco (Life Technologies, Paisley, UK). Sodium dodecyl sulphate (SDS) was purchased from BioShop (Burlington, ON, Canada), Coomassie brilliant blue was from Fluka (Buchs, Switzerland), and dithiothreitol (DTT) from Serva (Heidelberg, Germany). Redistilled phenol, sulphuric acid, isoamyl alcohol, and solvents were provided by POCH (Gliwice, Poland). All the chemicals used were of analytical grade.
Assessment of biological activity and determination of total phenolic content was conducted on 96-well transparent microplates (Nunclon, Nunc, Roskilde, Denmark) using an Elisa Reader Infinite Pro 200F (Tecan Group Ltd., Männedorf, Switzerland). Total sugar, glucans, and protein content were determined using a UV-VIS Evolution 300 spectrophotometer (Thermo Scientific, Lafayette, CO, USA).
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2

Insulin Aggregation Inhibition Assay

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Recombinant human insulin was purchased from SAFC bioscienses (Steinheim, Germany). Aloin (≥ 97%), Thioflavin T (approx. 65%), aloe-emodin, ascorbic acid, formic acid and TFA were purchased from Sigma-Aldrich (Stenheim, Germany). Sodium azide was purchased from Sigma-Aldrich (Switzerland). β-D-(+)-Glucose was purchased from Chromadex (USA). Disodium hydrogen phosphate, sodium chloride and acetonitrile were purchased from Merck Millipore (Darmstadt, Germany). Sodium dihydrogen phosphate, hydrochloric acid and sodium hydroxide were purchased from VWR Int. (Leuven, Belgium). Methanol (HPLC grade) was purchased from LAB-SCAN (Poland). Acetic acid was purchased from Applichem (Darmstadt, Germany). Water was in-house produced, 18 Ω, 0.22 μm.
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