Maxime rt premix oligo 18 primer kit

L-glutamic acid, Folin-Ciocalteu phenol reagent, sodium carbonate, gallic acid, aluminum chloride, sodium acetate, quercetin, 2,2′-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), N-Acetyl cysteine, and ascorbic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dimethyl sulfoxide (DMSO) was purchased from RCI Labscan (Bangkok, Thailand). Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), phosphate-buffered saline (PBS), and Hanks’ Balanced Salt solution (HBSS) were purchased from Thermo Scientific HyClone (Logan, UT, USA). Trypsin-EDTA (0.25%) was purchased from Gibco (Waltham, MA, USA). 3-(4,5-dimetylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) was purchased from Bio Basic (Markham, ON, Canada). Trizol was purchased from Invitrogen (Carlsbad, CA, USA). The maxime RT PreMix Oligo 18 Primer kit and RealMOD^TM Green W2 2x qPCR mix were purchased from iNtRON Biotechnology (Gyeonggi, South Korea). 2′, 7′-dichlorodihydrofluorescein diacetate (H₂DCFDA) was purchased from Molecular Probes (Eugene, OR, USA). The annexin V-Fluorescein isothiocyanate (FITC) apoptosis detection kit was purchased from BioLegend (San Diego, IL, USA).

HT-22 cells were treated with the designed treatments as described above. After that, total RNA was isolated using Trizol reagent (Invitrogen) according to the manufacturer’s instructions. The amount of the isolated RNA was measured with an absorbance at 260 nm using a NanoDrop spectrophotometer. Then, 1 µg of the isolated RNA was converted to complementary DNA (cDNA) using Maxime RT PreMix Oligo 18 Primer kit (iNtRON Biotechnology). The cDNA templates were amplified using RealMOD™ Green W2 2x qPCR mix (iNtRON Biotechnology) with the specific primers of SOD1, SOD2, CAT, GPx, and β-actin, following the previous study of Sukprasansap M et al. [19 (link)]. All PCRs were performed using an Exicycler^™ 96 (Bioneer). Quantitative gene expression was calculated by the 2^−ΔΔCT method with β-actin normalization.