Anti actb antibody

The fluorescence spectra were acquired on a SPEX Fluorolog 3-TCSPC spectrometer equipped with 1 cm path length cuvettes. The pH value was measured by PHS-3C Precision Ph/Mv Meter. The cell images were acquired on an Olympus Biological Microscope System. The 3 peptides, TAM-PEP (R4): TAM-RRRRNLWAAQRYGRELRRMSDKFVD, TAM-PEP (R6): TAM-RRRRRRNLWAAQRYG RELRRMSDKFVD and TAM-PEP (R8): TAM-RRRRRRRRNLWAAQRYGRELRRMSDKFVD, were synthesized by GL Biochem (Shanghai, China) Ltd. GO (XF-020, 50–200 nm) was purchased from XianFeng Nano Co. (Nanjing, China). The cDNA3.1-Bcl-xL plasmid was constructed by Genecreate Biological Engineering Co. (Wuhan, China) Tris-HCl, IPTG, PMSF, Anti-Bcl-xL antibody, Anti-ACTB antibody, HRP-conjugated Goat Anti-Rabbit IgG, Cell Counting Kit-8, Cytochrome C, BSA, RNase and lysozyme were purchased from Sangon Biotechnology Co. (Shanghai, China) BeyoECL Star and trypsin digestion solution were purchased from Beyotime Biotechnology Co. (Shanghai, China) Lipo ™ 2000 was purchased from Thermo Fisher Scientific Co.(Beijing, China)The deionized water was prepared on a Milli-Q water purification system and used throughout all experiments.

Expression of TNF-α, NF-κB p65, and TLR-4 was analyzed by Western blot. Eight rats per group were sacrificed by rapid decapitation at 48 h after surgery. The brains were removed, and the hippocampi were separated on ice. Part of each hippocampus was rapidly frozen in liquid nitrogen. The hippocampal samples were homogenized, and proteins were extracted by RIPA buffer. Protein concentrations were determined using a BCA protein assay kit (Thermo Fisher Scientific Inc., Waltham, MA, USA). The proteins were then separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) using 12% acrylamide gels and transferred onto a polyvinylidene difluoride (PVDF) membrane. The membrane was then incubated with the following primary antibodies at 4°C overnight: anti-TLR4 antibody (1:500, ab 13556, Abcam), anti-NF-κB p65 antibody (1:2,000, ab16502, Abcam), anti-TNF-α antibody (1:500, ab6671, Abcam). Anti-ACTB antibody (1:2,000, Sangon Biotech) was used as an internal control. After incubation with horseradish peroxidase (HRP)-conjugated goat anti-rabbit secondary antibody (1:2,000, ZB02301, ZSGB-BIO) for 2 h at room temperature, the blotted protein bands were visualized by enhanced chemiluminescence (ECL) Western blotting detection kit (P0018FS, Beyotime).