Incubation:Article Title: H19 lncRNA Promotes Skeletal Muscle Insulin Sensitivity in Part by Targeting AMPK
Article Snippet: Images were obtained under confocal microscopy (Leica SP5) at a magnification of 60×. .. For glutathione S-transferase (GST) pulldown, GST-AMPKα (4 μg, H00005563-P01; R&D Systems) or GST (202039; GenScript) was incubated with 8 μL of glutathione beads (16100; ThermoFisher) in 200 μL of binding buffer (BB; 25 mmol/L HEPES [pH 7.3], 100 mmol/L NaCl, 10% glycerol, 1% Triton X-100, 1 mmol/L dithiothreitol, 0.25 mmol/L phenylmethylsulfonyl fluoride, and 1× protease inhibitor cocktail [Calbiochem]) in the presence of 0.1% BSA at 4°C for 6 h. The beads were then washed with cold BB once, followed by incubation with 2 μg of FL-DUSP27 (TP314361; OriGene) in 200 μL of BB (without BSA) at 4°C overnight. ..
Article Title: H19 lncRNA Promotes Skeletal Muscle Insulin Sensitivity in Part by Targeting AMPK
Article Snippet: Images were obtained under confocal microscopy (Leica SP5) at a magnification of 60×. .. In Vitro Pulldown Assays For glutathione S-transferase (GST) pulldown, GST-AMPKα (4 μg, H00005563-P01; R&D Systems) or GST (202039; GenScript) was incubated with 8 μL of glutathione beads (16100; ThermoFisher) in 200 μL of binding buffer (BB; 25 mmol/L HEPES [pH 7.3], 100 mmol/L NaCl, 10% glycerol, 1% Triton X-100, 1 mmol/L dithiothreitol, 0.25 mmol/L phenylmethylsulfonyl fluoride, and 1× protease inhibitor cocktail [Calbiochem]) in the presence of 0.1% BSA at 4°C for 6 h. The beads were then washed with cold BB once, followed by incubation with 2 μg of FL-DUSP27 (TP314361; OriGene) in 200 μL of BB (without BSA) at 4°C overnight. ..
Binding Assay:Article Title: H19 lncRNA Promotes Skeletal Muscle Insulin Sensitivity in Part by Targeting AMPK
Article Snippet: Images were obtained under confocal microscopy (Leica SP5) at a magnification of 60×. .. For glutathione S-transferase (GST) pulldown, GST-AMPKα (4 μg, H00005563-P01; R&D Systems) or GST (202039; GenScript) was incubated with 8 μL of glutathione beads (16100; ThermoFisher) in 200 μL of binding buffer (BB; 25 mmol/L HEPES [pH 7.3], 100 mmol/L NaCl, 10% glycerol, 1% Triton X-100, 1 mmol/L dithiothreitol, 0.25 mmol/L phenylmethylsulfonyl fluoride, and 1× protease inhibitor cocktail [Calbiochem]) in the presence of 0.1% BSA at 4°C for 6 h. The beads were then washed with cold BB once, followed by incubation with 2 μg of FL-DUSP27 (TP314361; OriGene) in 200 μL of BB (without BSA) at 4°C overnight. ..
Article Title: H19 lncRNA Promotes Skeletal Muscle Insulin Sensitivity in Part by Targeting AMPK
Article Snippet: Images were obtained under confocal microscopy (Leica SP5) at a magnification of 60×. .. In Vitro Pulldown Assays For glutathione S-transferase (GST) pulldown, GST-AMPKα (4 μg, H00005563-P01; R&D Systems) or GST (202039; GenScript) was incubated with 8 μL of glutathione beads (16100; ThermoFisher) in 200 μL of binding buffer (BB; 25 mmol/L HEPES [pH 7.3], 100 mmol/L NaCl, 10% glycerol, 1% Triton X-100, 1 mmol/L dithiothreitol, 0.25 mmol/L phenylmethylsulfonyl fluoride, and 1× protease inhibitor cocktail [Calbiochem]) in the presence of 0.1% BSA at 4°C for 6 h. The beads were then washed with cold BB once, followed by incubation with 2 μg of FL-DUSP27 (TP314361; OriGene) in 200 μL of BB (without BSA) at 4°C overnight. ..
Protease Inhibitor:Article Title: H19 lncRNA Promotes Skeletal Muscle Insulin Sensitivity in Part by Targeting AMPK
Article Snippet: Images were obtained under confocal microscopy (Leica SP5) at a magnification of 60×. .. For glutathione S-transferase (GST) pulldown, GST-AMPKα (4 μg, H00005563-P01; R&D Systems) or GST (202039; GenScript) was incubated with 8 μL of glutathione beads (16100; ThermoFisher) in 200 μL of binding buffer (BB; 25 mmol/L HEPES [pH 7.3], 100 mmol/L NaCl, 10% glycerol, 1% Triton X-100, 1 mmol/L dithiothreitol, 0.25 mmol/L phenylmethylsulfonyl fluoride, and 1× protease inhibitor cocktail [Calbiochem]) in the presence of 0.1% BSA at 4°C for 6 h. The beads were then washed with cold BB once, followed by incubation with 2 μg of FL-DUSP27 (TP314361; OriGene) in 200 μL of BB (without BSA) at 4°C overnight. ..
Article Title: H19 lncRNA Promotes Skeletal Muscle Insulin Sensitivity in Part by Targeting AMPK
Article Snippet: Images were obtained under confocal microscopy (Leica SP5) at a magnification of 60×. .. In Vitro Pulldown Assays For glutathione S-transferase (GST) pulldown, GST-AMPKα (4 μg, H00005563-P01; R&D Systems) or GST (202039; GenScript) was incubated with 8 μL of glutathione beads (16100; ThermoFisher) in 200 μL of binding buffer (BB; 25 mmol/L HEPES [pH 7.3], 100 mmol/L NaCl, 10% glycerol, 1% Triton X-100, 1 mmol/L dithiothreitol, 0.25 mmol/L phenylmethylsulfonyl fluoride, and 1× protease inhibitor cocktail [Calbiochem]) in the presence of 0.1% BSA at 4°C for 6 h. The beads were then washed with cold BB once, followed by incubation with 2 μg of FL-DUSP27 (TP314361; OriGene) in 200 μL of BB (without BSA) at 4°C overnight. ..
In Vitro:Article Title: H19 lncRNA Promotes Skeletal Muscle Insulin Sensitivity in Part by Targeting AMPK
Article Snippet: Images were obtained under confocal microscopy (Leica SP5) at a magnification of 60×. .. In Vitro Pulldown Assays For glutathione S-transferase (GST) pulldown, GST-AMPKα (4 μg, H00005563-P01; R&D Systems) or GST (202039; GenScript) was incubated with 8 μL of glutathione beads (16100; ThermoFisher) in 200 μL of binding buffer (BB; 25 mmol/L HEPES [pH 7.3], 100 mmol/L NaCl, 10% glycerol, 1% Triton X-100, 1 mmol/L dithiothreitol, 0.25 mmol/L phenylmethylsulfonyl fluoride, and 1× protease inhibitor cocktail [Calbiochem]) in the presence of 0.1% BSA at 4°C for 6 h. The beads were then washed with cold BB once, followed by incubation with 2 μg of FL-DUSP27 (TP314361; OriGene) in 200 μL of BB (without BSA) at 4°C overnight. ..
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