Carbonyl cyanide

Manufactured by Merck Group

Sourced in United States

Carbonyl cyanide is a chemical compound used as a laboratory reagent. It functions as a proton ionophore, facilitating the transport of protons across cell membranes.

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Lab products found in correlation

7 protocols using carbonyl cyanide

Mitochondrial Respiration Modulation

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Oligomycin A, Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), rotenone, antimycin A, and 2-deoxy-d-glucose were purchased from Sigma-Aldrich. Oligomycin A and FCCP were dissolved in ethanol, antimycin A and rotenone were dissolved in DMSO before dilution. 2-deoxy-d-glucose was dissolved in supplemented XF base media and re-buffered to pH 7.4.

Lim T.K., Rone M.B., Lee S., Antel J.P, & Zhang J. (2015). Mitochondrial and bioenergetic dysfunction in trauma-induced painful peripheral neuropathy. Molecular Pain, 11, 58.

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Mitochondrial Function Assay Protocol

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Luciferin, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), pyruvate, palmitate, α-cyano-4-hydroxycinnamic acid (α-CCA), isoproterenol (ISO), blebbistatin, cyclosporin A (CsA), antimycin A, and genipin were from Sigma. Ethylene glycol tetraacetic acid (EGTA) was from Amresco. ABT-737 (ABT) was from Selleck Chemicals (Houston, Texas). Rotenone was from Calbiochem. MitoTEMPO was from Enzo Life Sciences. Mg²⁺-Green AM, Rhod-4 AM, BCECF AM, and mitoSOX were from Invitrogen (Eugene, Oregon). The tetrapeptide SS31 (D-arg-dmt-lys-phe-NH₂) was synthesized as described (Zhao et al., 2004 (link)).

Wang X., Zhang X., Wu D., Huang Z., Hou T., Jian C., Yu P., Lu F., Zhang R., Sun T., Li J., Qi W., Wang Y., Gao F, & Cheng H. (2017). Mitochondrial flashes regulate ATP homeostasis in the heart. eLife, 6, e23908.

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Mitochondrial Function Assay Protocol

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Triclosan (CAS 3380-34-5), analytical grade, ≥97.0% (HPLC), Dulbecco's phosphate buffered saline (PBS; Mg and Ca free), resazurin sodium salt (CAS 62758-13-8), carbonyl cyanide-(trifluoromethoxy) phenyl-hydrazone (FCCP, CAS 370-86-5), TPP⁺, malate and glutamate, MgATP, rotenone ≥95% (CAS 83-79-4), alamethicine (CAS 27061-78-5) and HEPES (CAS 7365-45-9) analytical grade ≥99.0% were from Sigma, 2,4 (α)-dinitrophenol (CAS 51-28-5) from Merck. The tissue culture media, RPMI 1460 glucose containing (15.8 mM) and RPMI1640 glucose free, trypsin 10× with EDTA, for cell detachment, were purchased from Lonza (Verviers, Belgium). For Complete medium, l-glutamine (Gibco 21875), heat inactivated 10% fetal bovine serum (FBS), PenStrep (penicillin 10,000 units and streptomycin 10,000 μg ml⁻¹), all from Gibco (Invitrogen, Carlsbad CA) were added. Glucose strips were from Lifescan (Johnson & Johnson, Espoo Finland). The fluorogenic dyes – JC-1 (5,5′, 6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolyl-carbocyanine iodide; dissolved in dimethyl sulfoxide DMSO), propidium iodide and calcein AM–were all obtained from Invitrogen (Carlsbad, CA, USA). The other chemicals were of analytical grade and purchased from local suppliers. Cultivation chamber slides # 154534 were from Labtek (Thermo Fisher Scientific, Vantaa Finland).

Ajao C., Andersson M.A., Teplova V.V., Nagy S., Gahmberg C.G., Andersson L.C., Hautaniemi M., Kakasi B., Roivainen M, & Salkinoja-Salonen M. (2015). Mitochondrial toxicity of triclosan on mammalian cells. Toxicology Reports, 2, 624-637.

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Dissecting Signaling Pathways in Cancer Cells

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Antibodies used: GAPDH (HyTest Ltd., clone 6C5, cat. # 5G4), caspase-3 (Abcam, ab4051), Cdk6 (Abcam, ab3126), Ras (Stressgen, now Enzo Life Sciences; ADI-KAP-GP001), p-ERK (Santa Cruz, sc-7383), Hsp70 (Abcam, ab181606), p27Kip1 (Cell Signaling, #2552), Nr2f1 (ABGENT, #AP14218a), tubulin (Merck, CP06), Cdk4 (NeoMarkers, MS-616-P1), Raf-1 (Santa Cruz Biotechnology, sc-133), Akt (Cell Signaling, #9272). Transfection reagents: jetPRIME (Polyplus, #114–15), polyethylenimine (PEI) (Polysciences, # 621405). Hsp90 inhibitors: geldanamycin (LC Laboratories, G-4500), PU-H71 (Tocris, #3104), pochoxime A (kindly provided by Nicolas Wissinger, University of Geneva). Other inhibitors: AZ628 (Tocris, #4836), PD98059 (Tocris, #1213), cycloheximide (Sigma, C7698), rapamycin (LC Laboratories, R-5000), aphidicolin (ACROS, BP615-1), hydroxyurea (fluorochem, #043351). All compounds were dissolved in DMSO. Growth factors: insulin (Sigma, I6634), recombinant human epidermal growth factor (EGF) (Lonza, CC-4017). Metabolic assay reagents: Oligomycin (SIGMA, 7535), carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) (SIGMA, C2920) and rotenone (SIGMA, R8875) with antimycin (SIGMA, A8674).

Echeverria P.C., Bhattacharya K., Joshi A., Wang T, & Picard D. (2019). The sensitivity to Hsp90 inhibitors of both normal and oncogenically transformed cells is determined by the equilibrium between cellular quiescence and activity. PLoS ONE, 14(2), e0208287.

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Mitochondrial Function and Cell Metabolism

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Carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP, Sigma-Aldrich, St. Louis, MO, USA), oligomycin A (Sigma-Aldrich, USA), 2-Deoxy-D-glucose (2DG, Sigma-Aldrich, USA), Trolox (Sigma-Aldrich, USA), ferrostatin (Sigma-Aldrich, USA) and bafilomycin (Sigma-Aldrich, USA) were used in the study.

Hendriks K.D., Joschko C.P., Hoogstra-Berends F., Heegsma J., Faber K.N, & Henning R.H. (2020). Hibernator-Derived Cells Show Superior Protection and Survival in Hypothermia Compared to Non-Hibernator Cells. International Journal of Molecular Sciences, 21(5), 1864.

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Systematic Evaluation of Cell Death Inducers

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Antimycin A (A8674), BAY87-2243 (SML2384), bupivacaine (B5274), carbonyl cyanide m-chlorophenylhydrazone (C2759), carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (C2920), chloroprocaine (1117008), dactinomycin (A1410), 2-deoxy glucose (D3179), 2,4-dinitrophenol (D198501), erastin (E7781), ferrostatin (SML0583), glucose (G7021), levobupivacaine (SML1092), lidocaine (L5647), mitoxantrone (M6545), necrostatin (N9037), oligomycin A (75351), prilocaine (P9547), ropivacaine (R0283), rotenone (557368), staurosporine (S6942), TNF-alpha (T6674), thapsigargin (T9033), tunicamycin (T7765) have been purchased from Merck-Sigma Aldrich. Oxaliplatin was purchased from Accord Healthcare (Ahmedabad, India). Bafilomycin A1 (1334), rapamycin (1292), torin 1 (4247) were purchased from Tocris bioscience (Bristol, UK). Smac-BV6 was purchased from CliniSciences (B4653), z-VAD-fmk (N1510.0025) was purchased from Bachem. Recombinant calreticulin was produced as described.8 14 (link)

Bezu L., Wu Chuang A., Sauvat A., Humeau J., Xie W., Cerrato G., Liu P., Zhao L., Zhang S., Le Naour J., Pol J., van Endert P., Kepp O., Barlesi F, & Kroemer G. (2022). Local anesthetics elicit immune-dependent anticancer effects. Journal for Immunotherapy of Cancer, 10(4), e004151.

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Evaluating Cellular Stress Responses

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The chemicals were obtained from the following sources: RPMI 1640, fetal bovine serum, L-glutamine, benzylpenicillin, streptomycin, Hanks’ balanced salt solution, trypsin, carbonyl cyanide p-(trifluoromethoxy)phenyl-hydrazone (FCCP), bisbenzimide (Hoechst 33342), propidium iodide (PI), 5, 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethylbenzimidazol carbocyanine iodide (JC-1), sodium palmitate, dichlorofluorescein-diacetate (DCFH-DA), alkaline phosphatase (from bovine intestinal mucosa), and albumin bovine serum (fatty acid and endotoxin free) from Sigma-Aldrich, St. Louis, MO, USA; Bcl-2 and GADD153 (F-168) antibodies from Santa Cruz Biotechnology, Dallas, TX, USA; phospho-IRE1α from Affinity BioReagents, Golden, CO, USA; phospho-eIF2α, cleaved caspase 3, and Bim from Cell Signaling, Danvers, MA, USA; and a horseradish-peroxidase-labeled donkey anti-rabbit or anti-mouse antibody from GE Healthcare, Uppsala, Sweden. All other chemicals of analytical grade were obtained from E. Merck, Darmstadt, Germany.

Wang X, & Welsh N. (2014). Bcl-2 maintains the mitochondrial membrane potential, but fails to affect production of reactive oxygen species and endoplasmic reticulum stress, in sodium palmitate-induced β-cell death. Upsala Journal of Medical Sciences, 119(4), 306-315.

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