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5 amino 4 imidazole carboxamide riboside aicar

Manufactured by LGC
Sourced in Denmark

5-Amino-4-imidazole carboxamide riboside (AICAR) is a nucleoside analog that functions as an AMP-activated protein kinase (AMPK) activator. It is a chemical compound used in research applications.

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2 protocols using 5 amino 4 imidazole carboxamide riboside aicar

1

Phosphorylation-specific Antibody Assay

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Primary antibodies specific for phospho-AMPKα (172), phospho-HER2 (1221/1222, 1248, 1196), phospho-HER2/phospho-EGFR (1248/1173), phospho-HER3 (1289), phospho-EGFR (1068, 1046/1047S, 1045), phospho-ACC (79), phospho-AMPK substrates (LXRXXpS/pT), AMPKα, AMPKβ1/2, ACC, PARP, HER2, EGFR, HER3, LKB1 and Myc-tag were purchased from Cell Signaling Technology. Anti-rabbit and anti-mouse secondary antibodies were also purchased from Cell Signaling Technology. Anti-phospho-EGFR (1142) was purchased from ECM Biosciences. Anti-Actin antibody was purchased from Sigma-Aldrich. 5-Amino-4-imidazole carboxamide riboside (AICAR), a pharmacological activator of AMPK (Toronto Research Chemicals, Toronto, ON) was dissolved in DMSO to make a 500mM stock concentration. EGF (Sigma) was prepared in a stock concentration of 100μg/ml in purified water. Compound C, an inhibitor of AMPK, was obtained from Calbiochem, and G418, used for selection of transfected cells, was obtained from Invitrogen. Control, AMPKα1/2, AMPKβ1, and AMPKβ2 siRNAs were purchased from Santa Cruz Biotechnology. All siRNA transfections were conducted in triplicate over a period of 72 hours using RNAiMax (Invitrogen) according to the manufacturer's recommendation.
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2

Measuring Cellular Lipid Metabolism

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To measure β-oxidation, the cells were incubated in the presence of [9,10-3 H(N)]-palmitic acid, and [ 3 H]-labelled water was measured as the product of NEFA oxidation (see the ESM Methods). Triacylglycerol (TAG) secretion, oleic acid (OA) and palmitic acid uptake, and incorporation of [ 14 C]OA and [ 14 C]glucose into TAG, were measured as described in the ESM Methods. Glucose uptake and production in response to insulin (100 nmol/l; Actrapid Penfill; Novo Nordisk, Bagsvaerd, Denmark) and glucose production in response to 5-amino-4-imidazole-carboxamideriboside (AICAR; 1 and 2 mmol/l; Toronto Research Chemicals, North York, ON, Canada) were assessed as previously described [6] .
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