A549 (adenocarcinoma cells) cell line, Dulbecco’s Modified Eagle’s Medium—high glucose (DMEM) power, Trypsin and 10× Phosphate buffer saline (PBS) Sterile, Carbonyl cyanide m-chlorophenyl hydrazone (CCCP) and Mitochondrial division inhibitor 1 (mdivi-1) were ordered from Merck Life Science, Science, Dorset, UK Limited. Cetuximab was bought from Medical Store (MS) Asanwa Ahmedabab Gujarat, India. Caffeine was ordered from CSPC Pharmaceutical Group Limited, Shijiazhuang, China. MitoTracker Green was bought from Thermo Fisher Scientific, Loughborough, UK. Fluorescence Nanodiamond (FND) was bought from FND BIOTECH, Inc. Taipei, Taiwan. PD153035 was bought from MedChemExpress LLC (MCE), Princeton, NJ, USA. Biotin EGF was bought from Molecular Probes, Invitrogen, Carlsbad, CA, USA. Both MitoTracker Green and mdivi-1 were dissolved as stock solutions in dimethyl sulfoxide (DMSO) for dilution in complete media.
Carbonyl cyanide m chlorophenyl hydrazone
Manufactured by Merck Group
Sourced in United States, Germany
Carbonyl cyanide m-chlorophenyl hydrazone is a chemical compound used in laboratory settings. It is a potent proton ionophore that uncouples oxidative phosphorylation in mitochondria. This compound is utilized in research applications to study cellular energy metabolism and related processes.
Automatically generated - may contain errors
Lab products found in correlation
Oligomycin, by Merck Group (10 mentions)
Rotenone, by Merck Group (9 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (7 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Ethylene glycol tetraacetic acid (egta), by Merck Group (5 mentions)
Valinomycin, by Merck Group (5 mentions)
Antimycin a, by Merck Group (5 mentions)
Bovine serum albumin (bsa), by Merck Group (5 mentions)
Sodium chloride (nacl), by Merck Group (5 mentions)
Ethidium bromide, by Merck Group (4 mentions)
Sucrose, by Merck Group (4 mentions)
Succinate, by Merck Group (4 mentions)
Kh2po4, by Merck Group (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Mdivi 1, by Merck Group (3 mentions)
Mitotracker green, by Thermo Fisher Scientific (3 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Glutamate, by Merck Group (3 mentions)
Malate, by Merck Group (3 mentions)
Ofloxacin, by Merck Group (3 mentions)
71 protocols using carbonyl cyanide m chlorophenyl hydrazone
1
Assessing Mitochondrial Dynamics in A549 Cells
2
Rat Renal Proximal Tubular Cells Protocol
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Rat renal proximal tubular cells (RTCs, NRK-52E) were purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan), and culture cells in Dulbecco’s modified Eagle’s medium (DMEM) were supplemented with an antibiotic/antifungal solution and 5% fetal bovine serum (FBS, pH 7.2). Cells were grown to 85%–95% confluence before they were placed in a humidified 37 °C incubator, and cells from passages 5–20 were used. FBS, DMEM, and tissue culture reagents were obtained from Invitrogen (Carlsbad, CA, USA). PFOS and 4-phenylbutyrate (4-PBA) were purchased from Sigma Chemical (St. Louis, MO, USA). Oligomycin, carbonyl cyanide m-chlorophenyl hydrazone (CCCP), and rotenone were obtained from Merck Millipore (Darmstadt, Germany); 3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was obtained from SERVA Electrophoresis (Berlin, Germany). l -Carnitine and U0126 were obtained from Healthmate (Taichung, Taiwan) and Tocris Cookson (Bristol, UK), respectively. Protein assay agents were purchased from Bio-Rad (Hercules, CA, USA). The chemical concentration and treatment duration for each assay were set in accordance with the protocols used in our previously published studies17 (link)–19 (link) or pilot studies.
3
Mitochondrial function evaluation protocol
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ADP, Alamethicin, 3-amino-1,2,4-triazole, P1,P5-di(adenosine-5′)pentaphosphate (Ap5A), Amplex Red, ATP, carbonyl cyanide m-chlorophenylhydrazone (CCCP), cyclosporine A (CsA), EGTA, fatty acid-free BSA, glucose, glucose-6-phosphate dehydrogenase, glutamate, hexokinase, horseradish peroxidase, malate, mannitol, MgCl2, NaCl, NADP, (NH4)2SO4, oligomycin, Phenol Red, phosphoenolpyruvate, pyruvate kinase, rotenone, succinate, sucrose, tert-butyl hydroperoxide and Tris were from Sigma-Aldrich (Saint Louis, MO, USA); Coomassie G-250 was from MP Biomedicals (Santa Ana, CA, USA); CaCl2, KCl, K2HPO4, KH2PO4 and Safranine O were from Merck (Darmstadt, Germany); Dihydroethidium, Mitotracker Green FM, Propidium Iodide and Sytox Green were from Life Technologies (Carlsbad, CA, USA). Other reagents of the highest purity available were from domestic suppliers. MitoQ, SkQ1 and SkQ3 were kindly provided by Dr. D.S. Esipov from the A.N. Belozersky Research Institute of Physico-Chemical Biology, MSU, Moscow, Russia.
4
PINK1 Expression and Mutagenesis Protocol
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Carbonyl cyanide m-chlorophenyl hydrazone (CCCP), epoxomicin and cycloheximide were purchased from Sigma-Aldrich, valinomycin from Axxora. siRNA transfections were performed with 20nM control (all stars negative control) or PINK1-specific siRNA (5′ GACGCTGTTCCTCGTTATGAA-3′) using HiPerfect (all from Qiagen) or Lipofectamine2000 (Invitrogen). siRNA resistant PINK1-V5 constructs have been described before [5 (link), 24 (link)]. The mutations p.F104A, p.L347P and p.I368N were introduced by site-directed mutagenesis and verified by Sanger sequencing. The standard transfection protocol for DNA was as follows: for one well of a 12-well plate 1 μg of DNA and 2.5 μl of Lipofectamine2000 (Invitrogen) were each mixed with 100 μl of Opti-MEM media (Invitrogen), incubated for 5 min at room temperature, mixed together and incubated 20 min before adding to the cells. For low-level expression DNA and lipofectamine amounts were reduced by 50–75% (total DNA per 12 well: 0.5 or 0.25 μg).
5
Intracellular ROS Quantification in Dictyostelium
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For the detection of intracellular ROS levels, 5 × 105 vegetative D. discoideum cells were settled on glass plates for 30 min. A 10 μM 2′,7′-Dichlorofluorescin diacetate (DCF-DA, Sigma-Aldrich) was added to the cells and allowed to incubate for 30 min. Images (1 frame/min) of cells have been taken with a Zeiss LSM800 microscope with and without the addition of 1 μM Carbonyl cyanide m-chlorophenylhydrazone (CCCP; Sigma- Aldrich) over a time period of 30 min. Alternatively, 5 × 105 vegetative WT and mutant cells, washed and resuspended in PB, were seeded into wells of a black 96-wells-plate. A 10 μM DCF-DA was added to the wells and left to incubate for 30 min. Measurements (every minute) were taken immediately after the addition of 1 μM CCCP to the respective wells for 30 min via the microplate reader Synergy HTX (BioTek). Background measurements were taken of HL5 media with 10 μM DCF-DA and subtracted from the data. Measurements have been conducted five times.
6
Autophagy Modulation in Cell Infection
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Carbonyl cyanide m-chlorophenyl hydrazone (CCCP), rapamycin, 3-methyladenine (3-MA), Mdivi-1, SBI-0206965 (ULK1 inhibitor), and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Cells were treated with various doses of the drugs prior to infection.
7
Radiolabeled Compounds for Transport Studies
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Inhibitors: 2-heptyl-4-hydroxyquinoline N-oxide (HQNO) and dicyclohexylcarbodiimide (DCCD), and ionophores: valinomycin, nigericin or carbonyl cyanide m-chlorophenyl hydrazone (CCCP) were from Sigma (St. Louis, MO). The following radiolabeled compounds were used: 109Cd (carrier-free) or sodium [U-14C]glutamate (7.4 GBq/mmol)—from Amersham, UK, 86RbCl (1.075 GBq/mmol), sodium [14C]benzoate (407 MBq/mmol), [3H]inulin (3.7 GBq/mmol) or [γ-32P]ATP (111 TBq/mmol)—from NEN™ Life Science Products (Boston, MA), while 32Pi—inorganic orthophosphate (740 MBq/mmol)—from the Institute of Nuclear Research, Świerk, Poland.
8
Cerebral Vessel Respiration Measurement
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The cerebral vessels were placed in ice‐cold respiration medium consisting of 0.5 mmol/L EGTA (Sigma, catalog No. E4378), 3 mmol/L MgCl2 6 hexahydrate (Sigma, catalog No. M9272), 60 mmol/L lactobionic acid (Sigma, catalog No. 153516), 20 mmol/L taurine (Sigma, catalog No. T0625), 10 mmol/L KH2PO4 (Sigma, catalog No. P5655), 20 mmol/L HEPES (ThermoFisher Scientific, catalog No. 15630080), 110 mmol/L D‐sucrose (Sigma, catalog No. S0389), and 1 g/L BSA at pH 7.1.27 High‐resolution oxygen consumption measurements were conducted in 2 mL of MiR05 using the Oroboros Oxygraph 2k (Oroboros Instruments, Innsbruck, Austria). Polarographic oxygen measurements were acquired at 2‐second intervals with the steady‐state rate of respiration calculated from a minimum of 30 data points and expressed as pmol s‐1 per mg wet weight. All respiration measurements were conducted at 37°C in a working range [O2] of ≈200 to 100 μmol/L. Respiration was measured with sequential titrations of: 1.25 mmol/L ADP (CalBiochem, catalog No. 117105), 2 mmol/L malate (Sigma, catalog No. M1000), 5 mmol/L pyruvate (Sigma, catalog No. P2256), 10 mmol/L glutamate (Sigma, catalog No. G1626), 10 mmol/L succinate (Sigma, catalog No. S2378), 0.5 μmol/L carbonyl cyanide m‐chlorophenyl hydrazone (Sigma, catalog No. C2759), 0.5 μmol/L rotenone (Sigma, catalog No. R8875), and 2.5 μmol/L antimycin‐A (Sigma, catalog No. A8674).
9
Apoptosis Signaling Pathway Assays
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Secondary mouse IgGκ BP-HRP antibody (sc-516102), mouse anti-Bax (sc-20067), mouse anti-Bcl-2 (sc-7382), mouse anti-caspase-9 (sc-56076), mouse anti-caspase-3 (sc-56053), mouse anti-MMP-9 (sc-93859), mouse anti-cytochrome C (sc-13561), mouse anti-LC3I/LC3II (sc-271625), mouse anti-p62/SQSTM1 (sc-28359), and mouse anti-β-actin (sc-69879) were purchased from Santa Cruz Biotechnology ((Santa Cruz, CA, USA). Dulbecco’s Modified Eagle Medium (DMEM) and Fetal Bovine Serum (SFB) were purchased from Gibco (GRAND Island, NE, USA). Bradford reagent was obtained from Bio-Rad. CellTiter-Glo® was purchased from Promega (WI, USA). Annexin V-FITC, camptothecin (CPT), dichlorodihydrofluorescein diacetate (H2DCFDA), Hoechst 33342, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), diphenyl-1-pyrenylphosphine (DPPP) DNase-free RNase, and LysoTracker® Red were purchased from Thermo Fisher Scientific (CA, USA). Carbonyl cyanide m-chlorophenylhydrazone (CCCP), glutathione (GSH), acridine orange (AO), tetramethylrhodamine ethyl ester (TMRE), propidium iodide (PI), o-phthaldialdehyde (OPA), and monodansylcadaverine (MDC) were purchased from Sigma-Aldrich (MO, USA).
10
Extraction and Characterization of Bioactive Compounds from Myrica gale
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Myrigalone A, myrigalone B and myrigalone D, and 2′,4′-dihydroxy-6′-methoxy-3′5′-dimethylchalcone (DMC) were extracted from Myrica gale fruits and plants as described [36 (link)] by Syngenta’s Jealott’s Hill International Research Centre (Bracknell, UK). Gibberellin A4+7 and paclobutrazol were purchased from Duchefa Biochemie (Haarlem, The Netherlands). Phloretin, dihydrochalcone, naringenin, neohesperidin dihydrochalcone, daphnetin, psoralen, angelicin, ferulic acid, acacetin, carbonyl cyanide m-chlorophenyl hydrazone (CCCP), 2,3,5-triiodobenzoic acid (TIBA), indole-3-acetic acid (IAA), aminoethoxyvinylglycine (AVG), L-kynurenine, and hydrogen peroxide solution (30% (w/w)) were purchased from Sigma-Aldrich (St Louis, MO, USA). 5-(4-Chlorophenyl)-4H-1,2,4-triazole-3-thiol (yucasin) was purchased from Carbosynth Ltd. (Compton, Berkshire, UK). 4-(2,4-dimethylphenyl)-2-(1H-indol-3-yl)-4-oxobutanoic acid (auxinole) was purchased from Cambridge Bioscience (Cambridge, UK). All the compounds were dissolved in DMSO except for Phloretin, dihydrochalcone, naringenin and neohesperidin dihydrochalcone which were dissolved in methanol. Controls were performed with basal solvent (0.1% (v/v) DMSO or methanol) as appropriate.
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