Ampkα d63g4 rabbit mab

Manufactured by Cell Signaling Technology

Sourced in United States

AMPKα (D63G4) rabbit mAb is a laboratory reagent used to detect and study the AMP-activated protein kinase (AMPK) alpha subunit. AMPK is a cellular energy sensor that plays a crucial role in regulating cellular metabolism. This monoclonal antibody can be used in various techniques to investigate AMPK and its related signaling pathways.

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3 protocols using ampkα d63g4 rabbit mab

Macrophage Polarization Modulation Protocol

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Tetraethylorthosilicate (TEOS), Na₂CO₃, ethanol, methanol, ammoniumhydroxide (NH₃·H₂O), hexadecyl-trimethyl-ammoniumbromide (CTAB) and potassium permanganate (KMnO₄) were purchased from SinopharmChemReagent Co., Ltd. (China). Metformin, lipopoly-saccharide (LPS) and Recombinant Murine IFN-γ were purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Murine IL-4 was provided by PeproTech Biotechnology Co., Ltd. (Suzhou, China). Polyclonal antibodies CD47, CD80 and CD206 were obtained from Proteintech Group, Inc. (Wuhan, China). Coumarin-6 was ordered from Aladdin Biochemical Technology Co., Ltd. (Shanghai, China). Mouse Tumor Necrosis Factor Alpha and Mouse IL-10 ELISA kit was provided by ABclonal Biotechnology Co., Ltd. (Wuhan, China). ELISA kits of Mouse iNOS and Mouse Arg-1 were obtained from Jonln Biotechnology Co., Ltd. (Shanghai, China). FITC-Anti-Mouse CD80 Antibody, FITC-Anti-Mouse CD206 and APC-Anti-Mouse CD206 Antibody were provided by Elabscience Biotechnology Co., Ltd. (Wuhan, China). β-Actin, AMPKα (D63G4) Rabbit mAb and Phospho-AMPKα (Thr172) (40H9) Rabbit mAb were ordered from Cell Signaling Technology, Inc. (MA, USA). DSPE-PEG-M2pep was purchased from SunLipo NanoTech Co., Ltd. (Shanghai, China).

Chong L., Jiang Y.W., Wang D., Chang P., Xu K, & Li J. (2023). Targeting and repolarizing M2-like tumor-associated macrophage-mediated MR imaging and tumor immunotherapy by biomimetic nanoparticles. Journal of Nanobiotechnology, 21, 401.

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Metformin and Compound C Mechanism

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Metformin hydrochloride (D150959) was purchased from Sigma-Aldrich (St Louis, MO, USA) and dissolved in sterile saline. Compound C (6-[4-(2-Piperidin-1-yl-ethoxy)-phenyl)]-3-pyridin-4-yl-pyyrazolo[1,5-a] pyrimidine) was obtained from Tocris (Cat. No. 3093) and dissolved in dimethyl sulfoxide and further diluted with sterile saline. Metformin and Compound C (10 mg/kg) were injected intraperitoneally. Phospho-Acetyl-CoA Carboxylase (Ser79) (3661, 1:500), Acetyl-CoA Carboxylase (C83B10) rabbit mAb (3676, 1:1000), AMPKα (D63G4) rabbit mAb (5832, 1:1000) and Phospho-AMPKα (Thr172) (40H9) rabbit mAb (2535, 1:1000) were obtained from Cell signaling technology (Beverly, MA, USA). The mouse monoclonal antibody β-actin (A1978, 1:5000) was the product of Sigma-Aldrich (St Louis, MO, USA). HRP-conjugated secondary antibodies for western blot were diluted as 1: 10,000, and were from Thermo Fisher Scientific (Waltham, MA, USA).

Ai H., Fang W., Hu H., Hu X, & Lu W. (2020). Antidiabetic Drug Metformin Ameliorates Depressive-Like Behavior in Mice with Chronic Restraint Stress via Activation of AMP-Activated Protein Kinase. Aging and Disease, 11(1), 31-43.

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Western Blot Analysis of Protein Biomarkers

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The cell lysates were homogenized in mammalian protein extraction buffer (Sigma). The resulting lysates were centrifuged at 12,000 rpm, and 4°C for 15 min, and the protein contents were measured using a protein-assay dye reagent concentrate (Bio-Rad Laboratories, Hercules, CA, USA). Subsequently, 10–15% sodium dodecyl sulphate-polyacrylamide gel electrophoresis was performed, followed by transfer onto nitrocellulose blotting membranes (Amersham, GE Healthcare Life Science, Germany). The antibodies used for immunoblotting are as follows: Bax rabbit mAb (1:1,000; Cell Signaling Technology), Bcl2 (D17C4) rabbit mAb (1:1,000; Cell Signaling Technology), inducible nitric oxide synthase (iNOS; NOS2) (1:500; Santa Cruz Biotechnology, Santa Cruz, CA, USA), AMPKα (D63G4) rabbit mAb (1:1,000; Cell Signaling Technology), Phospho-AMPKα (Thr172) (D4D6D) rabbit mAb (1:1,000; Cell Signaling Technology), caspase-3 rabbit mAb (1:1,000; Cell Signaling Technology), cleaved caspase-3 (Asp175) (5A1E) rabbit mAb (1:1,000; Cell Signaling Technology), PARP (46D11) rabbit mAb (1:1,000; Cell Signaling Technology), and β-actin (13E5) rabbit mAb (1:2,500; Cell Signaling Technology). The protein bands were visualized using an enhanced chemiluminescence method with an ELC kit (Millipore, Billerica, MA, USA) and were quantified using Quantity 1 v.4.6.7 (Bio-Rad).

Kim K., Kwak M.K., Bae G.D., Park E.Y., Baek D.J., Kim C.Y., Jang S.E., Jun H.S, & Oh Y.S. (2021). Allomyrina dichotoma larva extract attenuates free fatty acid-induced lipotoxicity in pancreatic beta cells. Nutrition Research and Practice, 15(3), 294-308.

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