Electrochemiluminescence ecl kit

APP-PS1-HEK293 cells were purchased from Shanghai Enzyme-linked Biotechnology (Shanghai, China). ICT (purity >98.3%) was obtained from Nanjing Zelang Medical Technologies (Nanjing, China). 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) was purchased from Biosharp (Hefei, China). A lactate dehydrogenase (LDH) assay kit was obtained from Beyotime (Shanghai, China).
An electrochemiluminescence (ECL) kit was purchased from Thermo Fisher (Waltham, MA, USA). A BACE1 Fluorescence Resonance Energy Transfer (FRET) Assay kit was obtained from Panvera (Madison, WI, USA). A PrimeScript™ RT Reagent kit and SYBR™ Premix Ex Taq were purchased from TaKaRa Biotechnology (Shiga, Japan). Antibodies against anti-BACE1, anti-presenilin 1, anti-AβP_1–40, anti-AβP_1–42, anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and goat anti-rabbit immunoglobulin (Ig)G H&L (DyLight 488) were obtained from Abcam (Cambridge, UK). Anti-C-terminal fragment β (CTFβ) antibody was purchased from Biolegend (San Diego, CA, USA). Anti-sAPPβ (6A1) antibody and anti-sAPPa (2B3) antibody were obtained from Immuno-Biological Laboratories (Fujioka-Shi, Japan). Horseradish peroxidase-conjugated goat anti-mouse or anti-rabbit IgG antibody was purchased from Abmart (Shanghai, China).

Dulbecco’s modified Eagle’s medium (DMEM) containing fetal bovine serum (FBS), penicillin‒streptomycin, and actinomycin D was purchased from Sigma-Aldrich (USA). TRIzol reagent, Lipofectamine 3000, anti-insulin-like growth factor 2 mRNA binding protein 3 (IGF2BP3) primary antibody, bicinchoninic acid (BCA) kit, electrochemiluminescence (ECL) kit, BODIPY-C11, and SYBR Premix Ex Taq were purchased from Thermo Fisher Scientific (USA). The CCK-8 reagent was purchased from Dojindo Laboratories (Kumamoto, Japan). EdU reagent and RIPA lysis buffer were purchased from Beyotime (Shanghai, China). Anti-solute carrier family 7 member 11 (SLC7A11), anti-m6A, anti-U2AF2, anti-IgG antibodies, and an iron assay kit were purchased from Abcam (Shanghai, China). A fluorescence in situ hybridization (FISH) kit was purchased from RiboBio (Guangzhou, China). The Magna MeRIP m6A kit was purchased from Millipore (USA). The Magnetic RNA Protein Pull-Down Kit was purchased from Pierce (USA).

The lung tissues were homogenized and protein concentration was determined using a Bradford reagent assay (Bio-Rad, Hercules, CA, USA). The proteins were separated via 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene fluoride (PVDF) membranes. The membranes were then incubated with 5% skim milk blocking solution for 1 h. After incubation, the membranes were blotted with primary antibody at 4 °C overnight. Primary antibodies used were as follows: COX2 (Abcam, Cambridge, UK), iNOS (Abcam), phospho (p)-IκBα (Cell Signaling, Danvers, MA, USA), IκBα (Cell Signaling), p-p65 (Abcam), p65 (Abcam), and β-actin (Cell Signaling). The membranes were incubated with horseradish peroxidase (HRP)-conjugated secondary goat immunoglobulin G (IgG; Jackson Immuno Research, West Grove, PA, USA) for 1 h. The bands were visualized using an electrochemiluminescence (ECL) kit (Thermo Fisher Scientific) and ChemiDoc (Bio-Rad). Quantitative analysis of protein expression was conducted using IMT i-Solution software (IMT i-Solution Inc., Vancouver, BC, Canada).