Atpenin a5

Manufactured by Cayman Chemical

Sourced in United Kingdom, Germany

Atpenin A5 is a chemical compound that functions as an inhibitor of the mitochondrial respiratory complex II. It is used in scientific research to study the role of complex II in cellular processes and metabolic pathways.

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Lab products found in correlation

7 protocols using atpenin a5

Metabolic Modulation in Immune Cells

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All reagents were from Sigma unless otherwise stated. Specific kits and linked reagents are described below. Non-canonical reagents are listed.
Rotenone, dehydroepiandrosterone (DHEA), thenoyltrifluoroacetone (TTFA), antimycin A, 2-deoxyglucose (2-DG), oligomycin, glutamate (pH 7.4 with NaOH), zymosan particles from Saccharomyces cerevisiae (including pHrodo, Thermo-Fisher), Yeast from Saccharomyces cerevisiae, LPS (lipopolysaccharide), luminol, DTAC (Dodecyltrimethylammonium chloride), Accutase, diazo-oxo-norleucine (DON), bis-phenylacetamido-thiadiazolyl-ethyl sulphide (BPTES), atpenin A5 (Cayman chemicals), dimethylmalonate, UK-5099, etomoxir, phorbol–myristate–acetate (PMA), mitotracker red CMX-Ros (Thermo-Fisher), all-trans retinoic acid, M-CSF (Peprotech), VAS-2870, Sotrastaurin (Cayman chemicals), N-Formylmethionyl-leucyl-phenylalanine (fMLF), Pam3CSK4 (Invivogen), uric acid crystals, R848 (Invivogen), 4′,6-Diamidino-2-Phenylindole (DAPI, Thermo-Fisher), Hoescht 33342 (Thermo-Fisher), 2-mercaptoethanol.

Davies L.C., Rice C.M., Palmieri E.M., Taylor P.R., Kuhns D.B, & McVicar D.W. (2017). Peritoneal tissue-resident macrophages are metabolically poised to engage microbes using tissue-niche fuels. Nature Communications, 8, 2074.

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Mitochondrial Respiration Assay Protocols

Cited 2 times

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S1QEL1.1, S1QEL2.1 [5 (link)], S1QEL1.719 [25 ] and S3QEL3 [6 (link)] were provided by Calico Life Sciences LLC (South San Francisco, CA) and AbbVie Inc. (Chicago, IL). They were maintained as 10 mM stocks in dimethylsulfoxide at room temperature, shielded from bright light and diluted in dimethylsulfoxide as required before use. Amplex UltraRed (Cat No. A36006) was from ThermoFisher; atpenin A5 (Cat No. 11898) and FCCP (Cat No. 15218) from Cayman Chemicals; piericidin A (Cat No. 2738-64-9) from Santa Cruz Biotechnology; and horseradish peroxidase (HRP) (Cat No. P8125), superoxide dismutase 1 (SOD1) (Cat No. S7571), rotenone (Cat No. R8875), myxothiazol (Cat No. T5580), succinate (Cat No. 14160), glycerol 3-phosphate (Cat No. 94124), malonate (Cat No. 792535) and glutamate (Cat No. 1294976) were from Sigma.

Gibbs ET I.I., Lerner C.A., Watson M.A., Wong H.S., Gerencser A.A, & Brand M.D. (2023). Site IQ in mitochondrial complex I generates S1QEL-sensitive superoxide/hydrogen peroxide in both the reverse and forward reactions. Biochemical Journal, 480(5), 363-384.

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Inhibition of Mitochondrial ETC Complexes in Osteoclastogenesis

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Mitochondrial ETC complex I inhibitor piericidin A (#15379) and complex II inhibitor atpenin A5 (#11898) were purchased from Cayman Chemical, while complex III inhibitor myxothiazol (T5580) was purchased from MilliporeSigma. Cultured BMM were treated with DMSO (control group), piericidin A (0.2 μM), atpenin A5 (0.4 μM), myxothiazol (0.2 μM), or antimycin (0.2 μM) along with methyl-pyruvate (1 mM) and uridine (400 μM) followed by RANKL stimulation. All concentrations were referred to published papers (42 (link), 43 (link)) and optimized for BMM treatment. TRAP staining was performed when mature OCs appeared in the control group.

Liu H., Zhai L., Liu Y., Lu D., Vander Ark A., Yang T, & Krawczyk C.M. (2023). The histone demethylase KDM5C controls female bone mass by promoting energy metabolism in osteoclasts. Science Advances, 9(14), eadg0731.

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Biochemical Assays for Oxidoreductase Activity

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Chemicals were purchased from Sigma: bovine serum albumin (BSA) essentially fatty acid free (#A6003), DDM (#D4641), hexaammineruthenium (III) chloride (#262005), horseradish peroxidase (#P8250), mannitol (#63559), NADH (#N8129), Q1 (#C7956), SOD (#S9697), sucrose (#S7903), and triphenyl-tetrazolium chloride (#T8877). 3–12% acrylamide gradient gel (#BN1001BOX), Amplex UltraRed (#A36006), NativeMark™ unstained protein standards (#LC0725), PageBlue Protein Staining Solution (#24620), and Pierce BCA protein assay kit (#23225) were from Thermo Fisher Scientific. Alamethicin (#11425) and atpenin A5 (#11898) were from Cayman Chemical.

Yoval-Sánchez B., Ansari F., James J., Niatsetskaya Z., Sosunov S., Filipenko P., Tikhonova I.G., Ten V., Wittig I., Rafikov R, & Galkin A. (2022). Redox-dependent loss of flavin by mitochondria complex I is different in brain and heart. Redox Biology, 51, 102258.

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Membrane Protein Purification and Characterization

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Most of the chemicals were purchased from Sigma, including mannitol (catalog no. 63559), sucrose (catalog no. 84097), essentially fatty acid–free bovine serum albumin (BSA; catalog no. A6003), NADH (catalog no. N8129), hexaammineruthenium (III) chloride (catalog no. 262005), Q₁ (catalog no. C7956), DDM (catalog no. S#D4641), and triphenyl-tetrazolium chloride (catalog no. T8877). Pierce bicinchoninic acid protein assay kit (catalog no. 23225), Amplex UltraRed (catalog no. A36006), horseradish peroxidase (catalog no. 012001), 3 to 12% acrylamide gradient gel, 8 × 8 cm (catalog no. BN1001BOX), NativeMark unstained protein standards (catalog no. LC0725), and PageBlue Protein Staining Solution (catalog no. 24620) were from Thermo Fisher Scientific. Alamethicin (catalog no. 11425) and atpenin A5 (catalog no. 11898) were from Cayman Chemical.

Ansari F., Yoval-Sánchez B., Niatsetskaya Z., Sosunov S., Stepanova A., Garcia C., Owusu-Ansah E., Ten V., Wittig I, & Galkin A. (2021). Quantification of NADH:ubiquinone oxidoreductase (complex I) content in biological samples. The Journal of Biological Chemistry, 297(4), 101204.

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Intracellular pH Acidification Protocols

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Pharmacological acidification of intracellular pH in C2C12 cells was achieved by acute inhibition of mitochondrial oxidative phosphorylation with 100 nM atpenin A5 (Cayman Chemical) and 0.5 mM antimycin A (Sigma Aldrich) for 1 h and 2 h, respectively. 100 μM monensin (Sigma Aldrich) were added 1h before administration of atpenin A5 or hypoxia to prevent cellular acidification. Intracellular pH acidification by extracellular equilibration was achieved as previously described (Nadtochiy et al., 2016). Briefly, cells were incubated with pH 6.4 medium supplemented with 30 mM NH₄Cl and 10 μM 5-(N-ethyl-N-isopropyl) amiloride (EIPA) for 15 min, followed by reincubation in pH 6.4 medium with 10 μM EIPA alone for 4 hours.

Reddy A., Bozi L.H., Yaghi O.K., Mills E.L., Xiao H., Nicholson H.E., Paschini M., Paulo J.A., Garrity R., Laznik-Bogoslavski D., Ferreira J.C., Carl C.S., Sjøberg K.A., Wojtaszewski J., Jeppesen J.F., Kiens B., Gygi S.P., Richter E.A., Mathis D, & Chouchani E.T. (2020). pH-Gated Succinate Secretion Regulates Muscle Remodeling in Response to Exercise. Cell, 183(1), 62-75.e17.

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Cellular Metabolism Profiling with Seahorse

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The cellular oxygen consumption rate (OCR) and the extracellular acidification rate (ECAR) were analyzed using a Seahorse 96 extracellular flux analyzer (Agilent, Waldbronn, Germany). THP-1 cells were plated in Seahorse 96-well cell culture plates one day prior to the measurements, and equilibrated for 30 min before recordings were made in Krebs Henseleit buffer (111 mM NaCl, 4.7 mM KCl, 1.25 mM CaCl₂, 2 mM MgSO₄, 1.2 mM Na₂HPO₄) supplemented with 5 mM l-glucose and 1 mM l-glutamine. Cells were treated with 1 μM rotenone (Sigma-Aldrich, Munich, Germany), 50 µM etomoxir (Cayman Chemicals), 30 µM Bis-2-(5-phenylacetamido-1,3,4-thiadiazol-2-yl)ethyl sulfide (BPTES, Sigma-Aldrich, Munich, Germany), 20 µM UK5099 (Sigma-Aldrich, Munich, Germany), or 1 µM atpenin A5 (Cayman Chemicals).

Fuhrmann D.C., Olesch C., Kurrle N., Schnütgen F., Zukunft S., Fleming I, & Brüne B. (2019). Chronic Hypoxia Enhances β-Oxidation-Dependent Electron Transport via Electron Transferring Flavoproteins. Cells, 8(2), 172.

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