β actin antibody 13e5

Shikonin, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Propidium Iodide (PI), FITC (fluorescein isothiocyanate)-labeled Annexin V, cholesterol, BCIP/NBT (5-bromo-4-chloro-3-indolyl-phosphate/nitro blue tetrazolium) substrate, and RIPA buffer were purchased from Sigma, St. Louis, MO. HBTU and DSPE-PEG (2000)-NH₂ were procured from Novabiochem and Avanti Polar Lipids, respectively. Unless otherwise stated, all chemicals, reagents, and organic solvent were purchased from local commercial suppliers and were used without further purification. Anti- receptor interacting protein (anti-RIP) antibody (ab72139) and Anti-RIP3 antibody (ab152130) were purchased from Abcam. β-actin antibody (13E5) was procured from Cell Signaling Technology. Goat anti-mouse IgG Alkaline Phosphatase (DC05L) and goat anti-rabbit IgG Alkaline Phosphatase (DC06L) were purchased from Calbiochem.

Western blotting was performed as previously described (Li et al., 2021 (link)). Briefly, 50 μg of proteins extracted from the hippocampus or prefrontal cortex was loaded onto SDS-PAGE and separated, then transferred to a nitrocellulose membrane. After blocking the nonspecific binding sites with skim milk, proteins were detected with the following antibodies. Antibodies specific for RBP-Jκ (#14613-1-AP, dilution 1:1,000) were provided by Proteintech, Inc. NF-κB p65 (#BD-PT5770, dilution 1:1,000) was provided by Biodragon. Notch1 (#AF5249, dilution 1:1,000), phospho-NF-κB p65 (Ser536) (#AF5881, dilution 1:1,000) and HES-1 (#AF2167, dilution 1:1,000) were purchased from Beyotime Biotechnology. COP1 (#bs-13925R, 1:1,000) antibody was purchased from Bioss Antibodies. β-actin antibody (13E5) (#4970S, 1:1,000) was purchased from Cell Signaling Technologies. Western blotting results were scanned by Odyssey CLx infrared fluorescence imaging system (LI-COR Biosciences). The relative expression levels of proteins were normalized according to their optical density and quantified in ImageJ software.