Beta tubulin rabbit polyclonal antibody

Total protein was extracted from tissues and cells and lysed in RIPA buffer (Beyotime, China) mixed with protease/phosphatase inhibitors (Sigma-Aldrich, USA). Samples were collected via centrifugation at 13,000 g and 4 °C for 5 min. The supernatants were boiled at 100 °C for 5 min in loading buffer. A BCA protein assay (Thermo Scientific, USA) was used to measure protein concentrations. Equal amounts of protein were separated using 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to polyvinylidene difluoride (PVDF) membranes (Millipore, USA). The membranes were incubated overnight at 4 °C with primary antibody (HIF-1α mouse monoclonal antibody, 1:2000, Novus; TNF alpha rabbit polyclonal antibody, 1:1000, Abcam; MCP-1 mouse monoclonal antibody, 1:1000, Novus; beta tubulin rabbit polyclonal antibody, 1:1000, Proteintech; GAPDH rabbit polyclonal antibody, 1:1000, Proteintech; beta actin rabbit polyclonal antibody, 1:1000, Proteintech). An Alexa Fluor 680/790-labeled goat anti-rabbit/goat anti-mouse IgG antibody (1:25,000, LI-COR Biosciences, USA) was used as the secondary antibody. An Odyssey infrared imaging system (LI-COR Biotechnology, USA) was employed to visualize the blots.

SFN was purchased from Aladdin (China). The CCK-8 assay kit was purchased from Dojindo (Japan). FITC Annexin V Apoptosis Detection Kit I was purchased from BD Biosciences (USA). DNase I was purchased from Sigma (USA). The TUNEL apoptosis detection kit and DAPI were purchased from Beyotime (China). DMEM/F12 medium, fetal bovine serum (FBS), 25% (w/v) trypsin-EDTA, Phosphate Buffered Saline (PBS), goat serum, Alexa fluor 594 labeled goat anti-chicken secondary antibodies and Alexa fluor 488 labeled goat anti-rabbit secondary antibodies, RIPA lysis buffer, protein phosphatase inhibitors, BCA protein assay kit, and the ECL reagent kit were purchased from Thermo Scientific (USA). Caspase-3 and cleaved Caspase-3 rabbit polyclonal antibodies, SB203580, p38 MAPK, and phospho-p38 MAPK rabbit mAb were purchased from Cell Signaling Technology (USA). GFAP chicken polyclonal antibody, GFAP rabbit polyclonal antibody, and AQP4 rabbit polyclonal antibody were purchased from Abcam (USA). Beta tubulin rabbit polyclonal antibody and horseradish peroxidase-conjugated goat anti-rabbit secondary antibodies were purchased from Proteintech (USA).
SFN was dissolved in dimethyl sulfoxide (DMSO) to a storage concentration of 100 mM. SB203580 was dissolved in dimethyl sulfoxide (DMSO) to a storage concentration of 10 mM. 0.01%(v/v) DMSO was used as vehicle.