RPMI tissue culture media, Lipofectamine 2000 were purchased from Invitrogen (Carlsbad, CA); Berberine (catalogue number B3251, purity ≥98%) was from Sigma (St. Louis, MO), Troglitazone, TRAIL were purchased from EMD Biosciences (Gibbstown, NJ). The antibodies utilized were obtained from the following sources: poly (ADP-ribose) polymerase (PARP), caspase-3, caspase 8, caspase 9, pAMPKT172, total AMPK, AMPKα1 and α2, DR4, DR5 from Cell Signaling Technologies (Danvers, MA), FLAG from Sigma (St. Louis, MO), GAPDH from Ambion Inc. (Austin, TX); TRIzol reagent from Invitrogen, (Carlsbad, CA), RT2 PCR Profiler PCR Array PAHS-012Z (Cat # 330231) was from Qiagen (Valencia, CA).
Ampkα1 and α2
Manufactured by Cell Signaling Technology
Sourced in United States
AMPKα1 and α2 are catalytic subunits of the AMP-activated protein kinase (AMPK) complex. AMPK is a sensor of cellular energy status that plays a key role in regulating metabolism. AMPKα1 and AMPKα2 are the two catalytic subunits that make up the AMPK complex.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Flag tag (flag), by Merck Group (1 mentions)
Berberine, by Merck Group (1 mentions)
Total ampk, by Cell Signaling Technology (1 mentions)
Caspase 8, by Cell Signaling Technology (1 mentions)
Caspase 9, by Cell Signaling Technology (1 mentions)
Pampkt172, by Cell Signaling Technology (1 mentions)
P akt ser473, by Cell Signaling Technology (1 mentions)
Phospho gsk 3β ser9, by Cell Signaling Technology (1 mentions)
Luciferase assay reagent, by Promega (1 mentions)
Compound c, by Merck Group (1 mentions)
Chir99021, by Merck Group (1 mentions)
Ly294002, by Merck Group (1 mentions)
Poly adp ribose polymerase parp, by Cell Signaling Technology (1 mentions)
Akt inhibitor 8, by Merck Group (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
β galactosidase assay kit, by Thermo Fisher Scientific (1 mentions)
Ar a014418, by Merck Group (1 mentions)
P gs ser641, by Cell Signaling Technology (1 mentions)
3 protocols using ampkα1 and α2
1
Berberine and TRAIL Synergistic Apoptosis
2
Signaling Pathways Modulation in Cells
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RPMI and DMEM, DMEM F12 tissue culture media, Lipofectamine 2000 and β-galactosidase assay kit were purchased from Invitrogen; the luciferase Assay Reagent from Promega (Madison, WI); Troglitazone, TRAIL and Cycloheximide (CHX) were purchased from EMD Biosciences (Gibbstown, NJ), Compound C, AKT Inhibitor VIII, LY294002, Kenpaullone and AR-A014418 were from EMD Millipore (Billerica, MA), CHIR 99021 was from Sigma (St. Louis, MO). The antibodies utilized were obtained from the following sources: poly (ADP-ribose) polymerase (PARP), caspase-3, GSK-3β, phospho-GSK-3βSer9, GSK3α, AKT, pAKTSer473, pAKT2Ser474, PPARγ, AMPKα1 and α2, GS, pGSSer641 from Cell Signaling Technologies (Danvers, MA); GAPDH from Ambion Inc. (Austin, TX). The GSK3β promoter luciferase plasmid (pGL3-GSK-3β-luc (−427/+66) was obtained from the laboratory of Dr. Daniel D. Billadeau, Mayo Clinic, Rochester, MN [34 (link)].
3
Western Blot Analysis of Protein Targets
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Protein (20 μg) in the soluble lysates were resolved by 12% SDS-PAGE, then transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore, USA). The membrane was blocked in 5% non-fat milk in PBS containing 0.1% Tween-20 (PBST) for 1 h at room temperature, and probed with antibodies specific to LC3 (Novus, USA), caspase-3 (Cell signaling), p62 (Cell signaling), eNOS (Cell signaling), p∼Ser1177-eNOS (Cell signaling), AMP-activated protein kinase (AMPKα1 and α2; Cell signaling) and p∼AMPK (α1 and α2; Cell signaling). Subsequently, the membrane was thoroughly washed in PBST, and incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies for 1 h at room temperature. Finally, the membrane was developed using the enhanced chemiluminescence detection method (Amersham, USA).
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