Oligomycin

Manufactured by Cayman Chemical

Sourced in United States

Oligomycin is a type of lab equipment used for research purposes. It functions as an inhibitor of the mitochondrial ATP synthase enzyme, which plays a crucial role in the production of cellular energy.

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Lab products found in correlation

Fluoro carbonyl cyanide phenylhydrazone (fccp), by Cayman Chemical (13 mentions) Rotenone, by Cayman Chemical (8 mentions) Rotenone, by Merck Group (8 mentions) Antimycin a, by Merck Group (8 mentions) Antimycin, by Merck Group (5 mentions) Antimycin a, by Cayman Chemical (4 mentions) D glucose, by Merck Group (4 mentions) Antimycin, by Cayman Chemical (4 mentions) Bafilomycin a1, by Merck Group (4 mentions) 2 deoxy d glucose, by Cayman Chemical (3 mentions) Seahorse xfe96 analyzer, by Agilent Technologies (3 mentions) L glutamine, by Merck Group (3 mentions) Adenosine diphosphate (adp), by Merck Group (3 mentions) Propidium iodide, by Thermo Fisher Scientific (3 mentions) Carbonyl cyanide m chlorophenyl hydrazine cccp, by Merck Group (2 mentions) Metformin, by Cayman Chemical (2 mentions) Methoxyamine hydrochloride, by Merck Group (2 mentions) Doxycycline hyclate, by Merck Group (2 mentions) Sodium pyruvate, by Merck Group (2 mentions) Rotenone antimycin a, by Cayman Chemical (2 mentions)

Close spelling variants of this product

Oligomycin A

42 protocols using oligomycin

Compound Screening for Drug Discovery

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Carbonyl cyanide m-chlorophenyl hydrazine (CCCP) and okadaic acid were from Sigma (Sigma-Aldrich, St. Louis, MO, USA). AZD1080, CHIR98014, cyclosporin A, GNE7915, LRRK2in1, oligomycin, rotenone, SB216763, sorafenib, staurosporine, and valinomycin were from Cayman Chemical Company (Ann Arbor, MI, USA). Tipranavir (#11285) was obtained through the NIH’s HIV Reagent Program, which is supported by National Institute of Allergy and Infectious Diseases. 10 mM stock solutions in dimethyl sulfoxide (DMSO) were prepared for all compounds with the exception of staurosporine (1mg/ml in ethyl acetate) and stored at −20°C until further use. Working dilutions were prepared fresh for each experiment.

, & Alavi M.V. (2021). Tau phosphorylation and OPA1 proteolysis are unrelated events: Implications for Alzheimer’s Disease.. Biochimica et biophysica acta. Molecular cell research, 1868(12), 119116.

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Neuronal Apoptosis Signaling Pathway

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The following chemicals were from Sigma Aldrich (St. Louis, MO): carbonyl cyanide m-chlorophenyl hydrazine (CCCP), forskolin, cycloheximide, and rotenone. Oligomycin, koningic acid, and GNE-3511 were from Cayman Chemicals (Ann Arbor, Ml). Hoescht 33342 was from ThermoFisher (Waltham, MA) and ethidiuim homodimer was from Biotium (Fremont, Ca). Antibodies used in this study are as follows: Anti-TUJ1 (BioLegend San Diego, CA; RRID: AB_10063408; WB: 1:5000; IF: 1:500), Anti-SCG10 (Shin et al., 2012b WB: 1:1000), anti-cleaved caspase 3 (Cell Signaling Technologies Danvers, MA RRID: AB_2341188; IF: 1:500). Nmnat2 was detected with a rabbit polyclonal antibody generated to a peptide corresponding to amino acids 232–253 from mouse NMNAT2 (YenZym Antibodies, South San Francisico, CA). Rabbit antiserum was purified on a sulfolink resin column coupled with the above-mentioned peptide (Thermo Fisher Scientific, Waltham MA). The glycine-eluted antibody was dialyzed in PBS and the specificity of the antibody confirmed by western immunoblotting using lysate DRGs lacking endogenous NMNAT2 upon CAS9/CRISPR knockout.

Summers D.W., Frey E., Walker L.J., Milbrandt J, & DiAntonio A. (2019). DLK activation synergizes with mitochondrial dysfunction to downregulate axon survival factors and promote SARM1-dependent axon degeneration. Molecular neurobiology, 57(2), 1146-1158.

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Mitochondrial Metabolism Regulation Assays

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Chemicals were purchased from the following sources: CP-91149, 2-deoxy-D-glucose (ApexBio, Houston, USA); antimycin A (Alfa Aesar, Tewksbury, USA); metformin, oligomycin, rotenone, FCCP, and NADP⁺ (Cayman Chemical, Ann Arbor, USA); MDIVI-1, mitochondrial fusion promoter M1, Glucose (HK) assay Kit (GAHK20), insulin (I9278), G6PDH antibody (A9521), NADPH, PAS staining kit (395B), and amyloglucosidase (A7420) (Sigma-Aldrich, St. Louis, USA); p-GYS, β-actin, DRP-1, MFN-1, AMPK, p-AMPK, PDH, and p-PDH antibodies were from Cell Signaling Technology (Danvers, USA), antibodies for GYS-2 (22371–1-AP), and PYGL (15851–1-AP) were from Proteintech Group (Rosemont, USA).

Wilcox P.L., Amerson H.V., Kuhlman E.G., Liu B.H., O'Malley D.M, & Sederoff R.R. (1996). Detection of a major gene for resistance to fusiform rust disease in loblolly pine by genomic mapping.. Proceedings of the National Academy of Sciences of the United States of America, 93(9), 3859-3864.

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Th17 Mitochondrial Respiration Profiling

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OCR and ECAR measurements were performed using a Seahorse XFe96
analyzer (Seahorse Biosciences). Seahorse XFe96 plates were coated with
0.56ug Cell-Tak (Corning) in 24 mL coating buffer (8 μL
H₂O+ 16 μL 0.1M NaHCO₃ (PH 8.0)) overnight at 4
°C, and washed 3x with H₂O before use. In
vitro cultured Th17 cells were collected at 96 h, washed twice
in Seahorse RPMI media PH7.4 (Agilent, 103576-100) and seeded onto the
coated plates at 100,000 per well. To measure the OCR and ECAR of KA-treated
cells, 30 μM KA (Santa Cruz) was added into Olfr2control Th17 cell samples at the beginning of the Seahorse measurement.
Respiratory rates were measured in RPMI Seahorse media, pH7.4 (Agilent)
supplemented with 10 mM glucose (Agilent) and 2 mM glutamine (Agilent) in
response to sequential injections of oligomycin (1 μM), FCCP (0.5
μM) and antimycin/rotenone (1 μM) (all Cayman Chemicals, Ann
Arbor, MI, USA).

Wu L., Hollinshead K.E., Hao Y., Au C., Kroehling L., Ng C., Lin W.Y., Li D., Silva H.M., Shin J., Lafaille J.J., Possemato R., Pacold M.E., Papagiannakopoulos T., Kimmelman A.C., Satija R, & Littman D.R. (2020). Niche-Selective Inhibition of Pathogenic Th17 Cells by Targeting Metabolic Redundancy. Cell, 182(3), 641-654.e20.

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Purchasing Reagents for Cell Culture Experiments

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All routine laboratory chemicals including salts, detergents, and buffers were purchased from Sigma-Aldrich (St Louis, MO). Special chemicals including oligomycin (11342), FCCP (15218), antimycin A (19433) were from Cayman Chemicals (Ann Arbor, Michigan). All tissue culture reagents were purchased from Thermo-Fisher Scientific with the following catalog numbers: F-12K medium (21127030); heat-inactivated fetal bovine serum (16140071); heat inactivated horse serum (26050088); G418 (10131027); puromycin (A1113802); Lipofectamine LTX (15338500).

Anderson C.J., Kahl A., Qian L., Stepanova A., Starkov A., Manfredi G., Iadecola C, & Zhou P. (2018). Prohibitin is a positive modulator of mitochondrial function in PC12 cells under oxidative stress. Journal of neurochemistry, 146(3), 235-250.

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Oligomycin Biochemical Assay Protocol

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Oligomycin was purchased from Cayman Chemical; Paraformaldehyde (16%) was obtained from Agar Scientific, and Propidium iodide from Thermo Fisher Scientific. Antimycin, CCCP, TMRM, Iodoacetate, Collagenase and Triton-x were all purchased from Sigma. All chemicals used were of analytical grade.

Tanton H., Voronina S., Evans A., Armstrong J., Sutton R., Criddle D.N., Haynes L., Schmid M.C., Campbell F., Costello E, & Tepikin A.V. (2018). F1F0-ATP Synthase Inhibitory Factor 1 in the Normal Pancreas and in Pancreatic Ductal Adenocarcinoma: Effects on Bioenergetics, Invasion and Proliferation. Frontiers in Physiology, 9, 833.

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Stable Isotope Tracer Metabolomics

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¹³C₃-labeled alanine (CLM-2184-H), ¹⁵N-labeled alanine (NLM-454), ¹³C₅-labeled glutamine (CLM-1822-H), ¹³C₃-labeled lactate (CLM-1579), ¹³C₃-labeled pyruvate (CLM-2440), and ¹³C_x,¹⁵N_x-labeled amino acid standard mix (MSK-A2) were acquired from Cambridge Isotope Laboratories. D-glucose (Sigma), DMSO (Sigma), doxycycline hyclate (Sigma), oligomycin (Cayman Chemicals), sodium pyruvate (Sigma), 2-ketobutyric acid sodium salt hydrate (Sigma), rotenone (Cayman Chemicals), FCCP (Cayman Chemicals), antimycin A (Cayman Chemicals), L-alanine (Sigma), L-alanine tert-butyl ester (Alfa Aesar), UK-5099 (Cayman Chemicals), methoxyamine hydrochloride (Sigma), and MTBSTA + 1% TBDMSCl (Sigma).

Parker S.J., Amendola C.R., Hollinshead K.E., Yu Q., Yamamoto K., Encarnación-Rosado J., Rose R.E., LaRue M.M., Sohn A.S., Biancur D.E., Paulo J.A., Gygi S.P., Jones D.R., Wang H., Philips M.R., Bar-Sagi D., Mancias J.D, & Kimmelman A.C. (2020). Selective alanine transporter utilization creates a targetable metabolic niche in pancreatic cancer. Cancer discovery, 10(7), 1018-1037.

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Metabolic Profiling of Organoids

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Extracellular acidification rate (ECAR) measurements were performed using an XFe96 analyzer (Agilent Technologies). Seahorse XFe96 plates (Agilent Technologies) were coated with a 10× dilution of BME in Dulbecco’s modified Eagle’s medium (DMEM; Sigma-Aldrich). IL-17A–treated or control organoids were collected with organoid harvest solution, washed once in DMEM basic medium, resuspended in DMEM basic medium with 2 mM glutamine, and seeded onto the coated plates at 175 μl per well. Respiratory rates were measured in response to sequential injections of glucose (10 mM), oligomycin (2 μM), and 2-DG (50 mM) (all Cayman Chemical). Values were normalized to a total number of cells in organoid culture well using the CyQUANT Cell Proliferation Assay (Thermo Fisher).

Konieczny P., Xing Y., Sidhu I., Subudhi I., Mansfield K.P., Hsieh B., Biancur D.E., Larsen S.B., Cammer M., Li D., Landén N.X., Loomis C., Heguy A., Tikhonova A.N., Tsirigos A, & Naik S. (2022). Interleukin-17 governs hypoxic adaptation of injured epithelium. Science (New York, N.Y.), 377(6602), eabg9302.

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Measuring Cellular Oxygen Consumption

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Oxygen consumption rates (OCR) were measured using a Seahorse XFe96 analyzer (Agilent) where cells were seeded into Seahorse XFe96 plates at 10,000 – 20,000 per well overnight depending on the cell line. Respiration in intact cells was measured in DMEM (Sigma #5030) supplemented with 10 mM glucose and 2 mM glutamine. Where indicated, cells were pre-treated with 2 μM Antimycin A 15 minutes prior to initial measurements. Respiratory rates were measured in response to sequential injections of oligomycin (2 μM), FCCP (0.5 μM) and rotenone / antimycin A (1 μM) (all Cayman Chemicals). Permeabilized cell respiratory rates were measured after cells were washed and resuspended in 1x MAS buffer containing either substrates pyruvate (10 mM) and malate (1 mM) or ascorbate and TMPD, along with ADP (4 mM) and XF PMP Reagent (1 nM) (Agilent Technologies). Phosphorylating (“State 3”) respiration was measured as described in Divakaruni et al. (2014) . Cells were immediately lysed in 25 μL/well Reagent A (Bio-Rad) after each assay, and protein quantified using the DC protein assay kit (Bio-Rad). OCR was normalized to micrograms total protein, as quantified by a standard curve.

Hollinshead K.E., Parker S.J., Eapen V.V., Encarnacion-Rosado J., Sohn A., Oncu T., Cammer M., Mancias J.D, & Kimmelman A.C. (2020). Respiratory Supercomplexes Promote Mitochondrial Efficiency and Growth in Severely Hypoxic Pancreatic Cancer. Cell reports, 33(1), 108231.

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Metabolomics Analyses Protocols

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Reagents used for metabolomics analyses included: methanol and ultra-pure water (LC-MS grade, EMD Millipore, Gibbstown, NJ); formic acid, acetic acid (Optima LC/MS grade; Fisher Chemical, Pittsburgh, PA); and butylated hydroxytoluene (BHT, TCI America; Portland, OR), as well as zirconium oxide beads (Next Advance; Averill Park, NY). Deuterium (d) labeled internal standards DHA-d₅, ARA-d₈, EPA-d₅, LA-d₄, and 9(S)-HODE-d₄ (Cayman Chemical, Ann Arbor, MI) were used for quantification of total and free fatty acids, and oxidized DHA derivatives, respectively. Sterile d₆-α-tocopherol emulsion (Fresenius-Kabi, Graz, Austria) and D-(+)-glucose (Sigma Aldrich, St. Louis MO) were used for embryo microinjection rescue studies. Reagents used for bioenergetics profiling included: oligomycin (Cayman Chemicals; Ann Arbor, MI), carbonyl cyanide 4-(trifluoromethoxy) phenyl-hydrazone (FCCP), and sodium azide (Sigma-Aldrich; St. Louis, MO).

McDougall M., Choi J., Kim H.K., Bobe G., Stevens J.F., Cadenas E., Tanguay R, & Traber M.G. (2017). Lethal Dysregulation of Energy Metabolism During Embryonic Vitamin E Deficiency. Free radical biology & medicine, 104, 324-332.

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