Oligomycin

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Oligomycin is a macrolide antibiotic that functions as an inhibitor of the mitochondrial ATP synthase (F1F0 ATPase) enzyme complex. It prevents the proton translocation necessary for ATP synthesis, thereby inhibiting oxidative phosphorylation.

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Lab products found in correlation

Antimycin, by Santa Cruz Biotechnology (3 mentions) Rotenone, by Santa Cruz Biotechnology (3 mentions) Antimycin a, by Santa Cruz Biotechnology (2 mentions) Adenosine diphosphate adp, by Merck Group (2 mentions) 6 diazo 5 oxo l norleucine don, by Merck Group (2 mentions) Doxycycline dox, by Merck Group (2 mentions) Fluoro carbonyl cyanide phenylhydrazone (fccp), by Santa Cruz Biotechnology (2 mentions) Sodium dichloroacetate dca, by Merck Group (2 mentions) Oxygraph o2k respirometer chamber, by Oroboros (1 mentions) Sodium oxamate, by Santa Cruz Biotechnology (1 mentions) Nb100 2220, by Novus Biologicals (1 mentions) Rabbit anti tomm20, by Santa Cruz Biotechnology (1 mentions) Mouse anti atp5b, by Santa Cruz Biotechnology (1 mentions) Gp62 c, by Novus Biologicals (1 mentions) Mouse anti parkin, by Cell Signaling Technology (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Mitotracker cmxros, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Bafilomycin a1, by Merck Group (1 mentions) Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)

7 protocols using oligomycin

Drug Treatments for Cell Growth Assays

Cited 1 time

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For 3-day growth competition experiments with drug-treated cells, we used the following drug concentrations: 2.5 μM sodium arsenite (Ricca Chemical, 714216); 2.5 μM oligomycin A (Santa Cruz Biotechnology, sc-201551); 50 nM rotenone (Sigma-Aldrich, R8875-1G); 10 μM CCCP (Cayman Chemicals, 25458); 5 μM BTdCPU (EMD Millipore, 324892); 10 nM thapsigargin (Sigma-Aldrich, T9033-.5MG); 100 nM tunicamycin (Calbiochem, 65438010); 1.25 μM EN6 (Sigma-Aldrich, SML2689-5MG)^{48 (link)}; 4 nM bafilomycin A1 (Selleck Chemicals, S1413); and 40 nM 17-DMAG (Selleck Chemicals, S1142). For overnight drug treatments, we used 5 μM sodium arsenite, 10 μM CCCP, 0.2 μM oligomycin, 5 μM antimycin A (Santa Cruz Biotechnology, sc-202467) or otherwise indicated in the figure legends. To inhibit the proteasome or autophagy, we used 2 μM carfilzomib (Selleck Chemicals, S2853) for 6 h or 700 nM bafilomycin A1 for 6 h, respectively. ISRIB (Sigma-Aldrich, SML0843) was used at a concentration of 200 nM.

Haakonsen D.L., Heider M., Ingersoll A.J., Vodehnal K., Witus S.R., Uenaka T., Wernig M, & Rapé M. (2024). Stress response silencing by an E3 ligase mutated in neurodegeneration. Nature, 626(8000), 874-880.

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Mitochondrial Respiration Profiling in Cells

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Cells were trypsinized, resuspended in 2 ml DMEM (2×10⁶ cells/ml), and transferred to an Oxygraph-O2K respirometer chamber (Oroboros Instruments, Innsbruck, Austria). The temperature was maintained at 37°C and O₂ set at a range of 100–200µM. Baseline respiratory flux was used to determine routine respiration; oligomycin (20 µM; Santa Cruz, # sc-201551A) was used to inhibit ATP synthase and measure leak rate. Carbonyl cyanide m-chlorophenylhydrazone (CCCP; 5 µM, Sigma #C-2759) was infused into the oxygraph chamber to determine uncoupled state. Cell O₂ consumption rates were calculated using DatLab software version 6 (Oroboros Instruments). Reserve respiratory capacity was calculated as the difference between uncoupled and routine respiration. Citrate synthase activity was determined spectrophotometrically (at 412 nm) in lysed cells using Ellman’s regent to determine the linear rate of thiol group (coenzyme A) produced by the condensation of oxaloacetate and acetyl-CoA (13 ).

Phillips C.M., Zatarain J.R., Nicholls M.E., Porter C., Widen S.G., Thanki K., Johnson P., Jawad M.U., Moyer M.P., Randall J.W., Hellmich J.L., Maskey M., Qiu S., Wood T.G., Druzhyna N., Szczesny B., Módis K., Szabo C., Chao C, & Hellmich M.R. (2017). Upregulation of Cystathionine-β-synthase in Colonic Epithelia Reprograms Metabolism and Promotes Carcinogenesis. Cancer research, 77(21), 5741-5754.

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Metabolic Modulation of Liver Cancer Cells

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The cells were treated with 2-DG (0–10 mM), sodium oxamate (0–50 mM), or oligomycin (0–1.0 µg/mL; Santa Cruz, Dallas, TX, USA) for 24 h to test major metabolic pathways that are utilized by HepG2 cells. Each chemical was dissolved in DMSO and further diluted to final concentrations. Cells were treated with E2 (Sigma-Aldrich, St. Louis, MO, USA), ERα agonist PPT (Fisher Scientific), or ERβ agonist DPN (Fisher Scientific) for 48 h to examine effects of E2 and ERs. E2 and ERs were dissolved to 1 μM in DMSO and diluted to 10 nM in culture medium. Vehicle DMSO was the control treatment. The concentration ranges of these chemicals are commonly used in liver cancer research and our previous studies [13 (link),14 (link),28 (link),29 (link),30 (link)].

Shen M., Xu M., Zhong F., Crist M.C., Prior A.B., Yang K., Allaire D.M., Choueiry F., Zhu J, & Shi H. (2021). A Multi-Omics Study Revealing the Metabolic Effects of Estrogen in Liver Cancer Cells HepG2. Cells, 10(2), 455.

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Mitochondrial Dysfunction and Autophagy Analysis

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Antimycin A (Santa Cruz), oligomycin (Santa Cruz), CCCP (Sigma), bafilomycin A1 (Sigma), and MitoTracker CMXROS (ThermoFisher) were resuspended in DMSO (Sigma). Antimycin A and oligomycin were stored in aliquots at −80°C. CCCP and bafilomycin A1 were stored in aliquots at −20°C. Primary antibodies for immunofluorescent staining include the following: rabbit anti‐LC3 (Sigma L7543, 1:5,000), rabbit anti‐TOMM20 (Santa Cruz sc‐11415, 1:1,000), mouse anti‐Parkin (Cell Signaling 4211, 1:1,000), mouse anti‐DNA (Millipore CBL186, 1:1,000), mouse anti‐ATP5B (Santa Cruz sc‐166462, 1:1,000), mouse anti‐PEX13 (Santa Cruz sc‐271477, 1:100), rabbit anti‐PMP70 (Thermo Scientific PA1‐650, 1:1,000), mouse anti‐Flag (Sigma 184‐200UG, 1:1,000), and rabbit anti‐WIPI2 (Abcam ab105459, 1:500). Secondary antibodies were conjugated to AlexaFluor488, AlexaFluor594, and/or AlexaFluor647 (Invitrogen, 1:750). Primary antibodies for Western blot analyses include the following: mouse anti‐PEX13 (Santa Cruz sc‐271477, 1:200), rabbit anti‐ATG7 (Sigma A2856, 1:1,000), rabbit anti‐FANCC (Fanconi Anemia Research Fund FANCC‐C2, 1:1,000), mouse anti‐SMURF1 (Sigma WH0057154M1, 1:1,000), guinea pig anti‐p62 (Progen GP62‐C, 1:1,000), rabbit anti‐LC3 (Novus NB100‐2220, 1:1,000), and HRP‐conjugated mouse anti‐actin (Santa Cruz sc‐47778‐HRP, 1:2,000).

Lee M.Y., Sumpter R J.r., Zou Z., Sirasanagandla S., Wei Y., Mishra P., Rosewich H., Crane D.I, & Levine B. (2016). Peroxisomal protein PEX13 functions in selective autophagy. EMBO Reports, 18(1), 48-60.

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Metabolic Modulation in Cancer Research

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The following drugs were used: 6‐Diazo‐5‐oxo‐l‐norleucine (Don, D2141, Sigma‐Aldrich); Sodium dichloroacetate (DCA, 347795, Sigma‐Aldrich); NCT‐502 and PHGDH inactive (19716 and 19717, Cayman); Doxycycline (Dox, D9891, Sigma‐Aldrich), Oligomycin (sc‐203342, Santa Cruz Biotechnology); FCCP (sc‐203578, Santa Cruz Biotechnology); Antimycin (sc‐202467, Santa Cruz Biotechnology); Rotenone (sc‐203242, Santa Cruz Biotechnology); Cyt.C (C2037, Sigma‐Aldrich) CB‐839 (10‐4556, Focus Biomolecules); Adenosine diphosphate (ADP, A2754, Sigma‐Aldrich).

Xu R., Jones W., Wilcz‐Villega E., Costa A.S., Rajeeve V., Bentham R.B., Bryson K., Nagano A., Yaman B., Olendo Barasa S., Wang Y., Chelala C., Cutillas P., Szabadkai G., Frezza C, & Bianchi K. (2020). The breast cancer oncogene IKKε coordinates mitochondrial function and serine metabolism. EMBO Reports, 21(9), e48260.

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Metabolic Modulation in Disease Models

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The following drugs were used: 6-Diazo-5-oxo-L-norleucine (Don, D2141, Sigma Aldrich); Sodium dichloroacetate (DCA, 347795, Sigma Aldrich); NCT-502 and PHGDH inactive (19716 and 19717, Cayman); Doxycycline (Dox, D9891, Sigma Aldrich), Oligomycin (sc-203342, Santa Cruz Biotechnology); FCCP (sc-203578, Santa Cruz Biotechnology); Antimycin (sc-202467, Santa Cruz Biotechnology); Rotenone (sc-203242, Santa Cruz Biotechnology);Cyt.C (C2037, Sigma Aldrich) CB-839 (10-4556, Focus Biomolecules); Adenosine diphosphate (ADP, A2754, Sigma Aldrich).

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Mitochondrial Respiration Assay

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Culture media and supplements were purchased from Thermo Scientific, except DMEM without glucose (cat. D5030), that was from Sigma-Aldrich. Oligomycin, CCCP, rotenone, and antimycin A were from Santa Cruz Biotechnology. All the other reagents were purchased from Sigma-Aldrich, unless stated differently.

Kakimoto P.A., Zorzano A., & Kowaltowski A.J. (2020). Increased glycolysis is an early outcome of palmitate-mediated lipotoxicity.

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