Cells or tissues were lysed in RIPA buffer containing protease and phosphatase inhibitor cocktail. After the protein concentration was normalized by BCA Protein Quantitative Kit, the samples were separated by SDS-PAGE, immunoblotted with the indicated antibodies. Anti-SOX8 (1:1000, 20627-1-AP), anti-15-LOX (1:1000, 13073-1-AP), anti-PKM1(1:1000, 15821-1-AP), anti-NRF2 (1:1000, 66504-1-Ig), anti-Transferrin (1:1000 17435-1-AP), anti-GPX4 (1:1000 67763-1-Ig), anti-ACLY (1:1000 15421-1-AP), anti-GAPDH (1:1000 60004-1-Ig), anti-TOM20 (1:1000 11802-1-AP), anti-TIM23 (1:1000 11123-1-AP), anti-G6PD (1:1000 66373-1-Ig), anti-INSIG1 (1:1000, 24766-1-AP), anti-INSIG2 (1:1000, 24766-1-AP), anti-DDK (1:1000, 66008-3-Ig), anti-AIF(1:1000, 17984-1-AP), anti-LPCAT3 (1:1000, 67882-1-AP) and anti-SLC7A11 (1:1000, 26864-1-AP) were purchased from Proteintech (Rosemount, IL, USA). Anti-SCD1(1:1000, 2794), anti-pACC (1:1000, 29119-1-AP), anti-ACC (1:1000, 3662), anti-FASN (1:1000, 3180), anti-Cleaved Caspase3 (1:1000, 9664), anti-LC3B (1:1000, 3868) were got from CST (Danvers, MA, USA). Anti-SREBP1 (1:1000, sc-13551) was from Santa Cruz. Anti-Transferrin Receptor (1:1000, ab214039), anti-pP53 (1:1000, ab1431), anti-AMPKα2 (1:1000, ab214425), anti-pAMPK (1:1000, ab133448), anti-ACSL4 (1:1000, ab155282) was from Abcam.
Anti ampkα
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-AMPKα is a lab equipment product that specifically targets and binds to the AMP-activated protein kinase alpha (AMPKα) subunit. AMPKα is a catalytic subunit of the AMPK complex, which plays a crucial role in cellular energy homeostasis. The primary function of Anti-AMPKα is to facilitate the detection, quantification, and study of AMPKα in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Aicar, by Merck Group (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Penicillin, by Merck Group (2 mentions)
Anti ampk, by Santa Cruz Biotechnology (2 mentions)
Anti β actin, by Santa Cruz Biotechnology (2 mentions)
Anti phospho akt, by Santa Cruz Biotechnology (2 mentions)
Anti lox 1, by Santa Cruz Biotechnology (2 mentions)
Anti srebp 1, by Santa Cruz Biotechnology (1 mentions)
Anti slc7a11, by Proteintech (1 mentions)
Anti p p53, by Santa Cruz Biotechnology (1 mentions)
Anti aif, by Proteintech (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Cell Signaling Technology (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Rhodamine 123, by Merck Group (1 mentions)
Anti nitrotyrosine nt, by Merck Group (1 mentions)
Anti β actin antibody, by Merck Group (1 mentions)
Phosphorylated acetyl coa carboxylase, by Cell Signaling Technology (1 mentions)
Anti 4 hydroxynonenal 4 hne, by Merck Group (1 mentions)
B cell lymphoma 2, by Cell Signaling Technology (1 mentions)
5 protocols using anti ampkα
1
Comprehensive Protein Expression Analysis
2
Compound C Inhibits AMPK-Mediated Signaling
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AA, compound C, and STO-609 were purchased from Calbiochem (San Diego, CA, USA). Antibodies directed poly(ADP-ribose)polymerase (PARP), caspase-3, B cell lymphoma- (Bcl-) 2, phosphorylated AMPKα, phosphorylated acetyl-CoA carboxylase (ACC), ACC, phosphorylated Ca2+/calmodulin-dependent kinase kinase 2 (CaMKK2), liver kinase B1 (LKB1), and horseradish peroxidase-conjugated secondary antibodies were obtained from Cell Signaling Technology (Beverly, MA, USA). Anti-AMPKα and anti-CaMKK2 antibodies were supplied from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-nitrotyrosine (NT) and anti-4-hydroxynonenal (4-HNE) antibodies were purchased from Millipore (Temecula, CA, USA) and Abcam (Cambridge, UK), respectively. Fluo-4-acetoxymethyl ester (Fluo-4) was obtained from Invitrogen (Carlsbad, CA, USA). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), rhodamine 123, 2′,7′-dichlorofluorescein diacetate (DCFH-DA), CCl4, anti-β-actin antibody, and other reagents were supplied from Sigma-Aldrich (St. Louis, MO, USA).
3
HUVEC Culture and Characterization
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Human umbilical vein endothelial cells (HUVECs) were obtained from ATCC. HUVECs were cultured with M199 basal medium supplemented with low-serum growth supplement and penicillin (50 IU/ml)-streptomycin (50 μg/ml). trypsin-EDTA was used to passage cells. M199 and trypsin-EDTA were obtained from Gibco (Grand Island, NY, USA). Low-serum growth supplement was purchased from Cascade (Portland, OR, USA). Additionally, 5,58,6,68-tetraethylbenzimidazolcarbocyanine iodide (JC-1) were purchased from BioVision (Palo Alto, CA, USA). LY294002, dihydroethidium (DHE), Apocynin, 1,2-bis(o-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA), DCHC, AICAR, sodium nitroprusside (SNP), Ro-320432, penicillin and streptomycin were all purchased from Sigma (St. Louis, MO, USA). Anti-β-actin, anti-AMPK, anti-AMPK-α, anti-AKT, anti-phospho AKT, anti-LOX-1, anti-SIRT1, anti-eNOS were all obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). HRP-conjugated anti-rabbit secondary antibodies were purchased from Transduction Laboratories (CA, USA). (TUNEL) staining kit was obtained from Boehringer Mannheim (Mannheim, Germany) was purchased from Selleckchem. Fura-2 AM and the EnzChek caspase 3 assay kit were purchased from Molecular Probes (Eugene, OR). SOD activity assay kit was obtained from Calbiochem (San Diego, CA). eNOS kit was purchased from R&D Systems (Minneapolis, MN, USA).
4
Calcium-Dependent Regulation of AMPK Signaling
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Alexa Fluor 488 donkey anti-rabbit IgG (A21206) antibody was from Life Technologies Corp; fetal bovine serum (16000-044), Dulbecco’s Modified Eagle medium (DMEM; 11995-040) and RPMI 1640 medium (11875-093) from Gibco. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT; M2128), pepstatin A (P5318), 3-Methyladenine (3-MA; M9281) and anti-LC3B antibody (L7543) from Sigma; BAPTA/AM (196419) and compound C (171260) from Calbiochem; E64d (4545) and STO-609 (1551) form Tocris Bioscience; anti-TRPC5 (ACC-020) form Alomone labs; anti-ATCB (sc-47778) from Santa Cruz Biotechnology; anti-AMPKα (#2532 S), anti-phospho-AMPKα (Thr172) (#2535), anti-mTOR (#2972), anti-phospho-mTOR (Ser2448) (#2971), anti-CaMKKβ (#4436), anti-phospho-CaMKKβ (Thr286) (#12716), anti-phospho-p70S6K (Thr389) (#9205), anti-p70S6K (#9202) from Cell Signaling Technology.
5
Endothelial Cell Culture and Molecular Characterization
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Human umbilical vein endothelial cells (HUVECs) were obtained from ATCC. HUVECs were cultured with M199 basal medium supplemented with low-serum growth supplement and penicillin (50 IU/ml)-streptomycin (50 μg/ml). trypsin-EDTA was used to passage cells. M199 and trypsin-EDTA were obtained from Gibco (Grand Island, NY, USA). Low-serum growth supplement was purchased from Cascade (Portland, OR, USA). Additionally. Baicalein, AICAR, Compound C, Apocynin, penicillin and streptomycin were all purchased from Sigma (St. Louis, MO, USA). Anti-β-actin, anti-AMPK, anti-AMPK-α, anti-phospho AKT, anti-LOX-1, anti-PKCα, anti-PKCβ, anti-PKCγ, anti-PKCδ, I-κB, NF-κBp65, anti-Cu, Zn SOD, anti-Mn SOD were all obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
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