Phospho p38 mapk thr180 tyr182 antibody 9211

Protein extraction was performed using protocols described previously [33 (link)]. The concentrations of the total proteins were measured with a Bradford Protein Assay Kit (Beijing Solarbio Science & Technology Co., Ltd.), and 0.05 mg of each sample was separated by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were further transferred to Immobilon-P transfer membranes (Millipore, Billerica, USA), then phosphorylation of FgHog1 and total FgHog1 were detected by Phospho-p38 MAPK (Thr180/Tyr182) Antibody #9211 (Cell Signaling Technology Inc., Boston, MA, USA) and anti-Hog1 antibody sc165978 (Santa Cruz, CA, USA), respectively. Protein abundance was quantified with ImageJ software. The experiments were performed three times independently.

Most of the material used in this study was purchased as we reported earlier [4 (link),53 (link),68 (link),96 (link),97 (link),98 (link),99 (link),100 (link)]. Aβ 1–40 (AS-24235), Aβ 1–42 (AS-20276), Aβ 1–28 (AS-24231), Aβ 25–35 (AS-24227), and Aβ 1–16 (AS-24225) were purchased from AnaSpec. Biotin-HN (HN, B-018--26, UniProt Q8IVG9) was purchased from Phoenix Pharmaceuticals. Phosphate Buffered Saline (PBS), nitrocellulose membranes, streptavidin–horseradish peroxidase (HRP) conjugate, Ponceau S solution, chelerythrine chloride, and ACh were purchased from Sigma-Aldrich. Mouse α-tubulin monoclonal antibody (DM1A), goat anti-rabbit IgG (H+L) secondary antibody (HRP, 31466), 3,3′,5,5′-tetramethylbenzidine (TMB), BCA protein assay kit, super signal west pico luminol (chemiluminescence) reagent, lipofectamine 2000 transfection reagent, and the Halt protease and phosphatase inhibitor cocktail were from ThermoFisher. Donkey anti-mouse IgG (HRP) (ab205724) was purchased from Abcam. m-IgGκ BP-HRP was obtained from Santa Cruz Biotechnology. SignalSilence p53 siRNA I (6231), SignalSilence control siRNA (Unconjugated, 6568), rabbit p53 antibody (9282), p38 MAPK antibody (9212) that detects endogenous levels of total p38α/β/γ MAPK, phospho-p38 MAPK (Thr180/Tyr182) antibody (9211), PKCα antibody (2056), and SB203580 (5633S) were purchased from Cell Signaling Technology (Danvers, MA, USA).