Membrane protein extraction kit p0033

Rat hippocampus and cortex were isolated and homogenized in 10 volumes (ml/g wet tissue) homogenate buffer containing 50 mM Tris-HCl, pH 7.0, 0.5 mM PMSF, 2.5 mM EDTA, 2.5 mM EGTA, 2.0 mM Na₃VO₄,100 mM NaF and 1:1000 protease inhibitor cocktail (Sigma-Aldrich, St. Louis, MO, USA). Then the homogenates were sonicated and stored at −80°C for Western blotting. The membrane proteins were extracted by using the membrane protein extraction kit P0033 from Beyotime (Shanghai, China) according to the manufacturer's instruction.

Dopamine hydrochloride, 1,3,5-Trimethylbenzene (TMB) (97 %, A.R.)and pluronic F127 (98 %, A.R.) were bought from Aladdin Chemistry Co. KMnO₄ (≥99 %, A.R.) were bought from Sinopharm Chemical Reagent Co. Fibroblast activation protein antibody (FAP1) (AF5344, 50 μL) and α-smooth muscle actin antibody (α-SMA) (BF9212, 50 μL) were purchased from Affinity. Recombinant mouse TGF-β1 protein (7666-MB-005/CF, 5 μg) was purchased from RD systems. Ultra-Low adhesion (ULA) U-bottom 96-well plates (174925) were purchased from Thermofisher Scientific Inc. HSP90 antibody (AH732, 40 μL) and membrane protein extraction kit (P0033, 100 T) were bought from Beyotime Biotechnology Co. Calcein-AM/Propidium Iodide Live/Dead Cell Double Staining Kit (CA1630, 500 T) was bought from Beijing Solarbio Science & Technology Co.