Amyloid β1 40 human elisa kit

After the homogenization in RIPA buffer containing a cocktail of protease inhibitors, the harvested brain homogenates were completely lysed under constant 1 h agitation and then centrifuged at 10,000 × g (Biofuge stratos, Thermo Fisher, Germany) for 20 min at 4 °C. The supernatants were collected as soluble fractions. The centrifugal precipitates were further sonicated in 70% (w/v) formic acid, followed by centrifugation at 10,000 × g (Biofuge stratos, Thermo Fisher, Germany) for 30 min at 4 °C. The supernatants neutralized by 1 mol/L Tris Buffer were retrieved as insoluble fractions. Thereafter, the levels of Aβ_1‒40 and Aβ_1‒42 in soluble and insoluble fractions were determined in accordance with the operating instructions of Amyloid-β1–40 Human ELISA Kit and Amyloid-β1–42 Human ELISA Kit (Elabscience, China), respectively.

Rapamycin (Rapa), coumarin-6 (Cou-6), and okadaic acid (OA) were purchased from Sigma–Aldrich Inc. (St.Louis, MO, USA). Gypenoside XVII (GP-17) was attained from Push Biotechnology Co., Ltd. (Chengdu, China). RP-1 (CAPDTKTQ) and Aβ_1–42 were synthesized by Guotai Biotechnology Co., Ltd. (Hefei, China) and Chinese Peptide Co., Ltd. (Hangzhou, China), respectively. 1,1′-Dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI), 3,3′-dioctadecyloxacarbocyanine perchlorate (DiO), Cell Counting Kit-8 (CCK-8), Total ROS assay kit and BCA Protein Assay Kit were purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Amyloid-β1–40 Human ELISA Kit and Amyloid-β1–42 Human ELISA Kit were purchased from Elabscience Biotechnology Co., Ltd. (Shanghai, China). Lysotracker Red DND-99 and Lysosenser Yellow/Blue dextran were purchased from Thermo Fisher Scientific, Inc. (Eugene, OR, USA). The mouse brain microvascular endothelial cells (bEnd.3), mouse hippocampal neuronal cells (HT22), and murine neonatal microglial cells (BV2) were obtained from the Chinese Academy of Sciences Cell Bank (Shanghai, China). All chemicals used were analytical or reagent grade.

Rapamycin (Rapa), coumarin-6 (Cou-6), and okadaic acid (OA) were purchased from Sigma–Aldrich Inc. (St.Louis, MO, USA). Gypenoside XVII (GP-17) was attained from Push Biotechnology Co., Ltd. (Chengdu, China). RP-1 (CAPDTKTQ) and Aβ_1–42 were synthesized by Guotai Biotechnology Co., Ltd. (Hefei, China) and Chinese Peptide Co., Ltd. (Hangzhou, China), respectively. 1,1′-Dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI), 3,3′-dioctadecyloxacarbocyanine perchlorate (DiO), Cell Counting Kit-8 (CCK-8), Total ROS assay kit and BCA Protein Assay Kit were purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Amyloid-β1–40 Human ELISA Kit and Amyloid-β1–42 Human ELISA Kit were purchased from Elabscience Biotechnology Co., Ltd. (Shanghai, China). Lysotracker Red DND-99 and Lysosenser Yellow/Blue dextran were purchased from Thermo Fisher Scientific, Inc. (Eugene, OR, USA). The mouse brain microvascular endothelial cells (bEnd.3), mouse hippocampal neuronal cells (HT22), and murine neonatal microglial cells (BV2) were obtained from the Chinese Academy of Sciences Cell Bank (Shanghai, China). All chemicals used were analytical or reagent grade.