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CYTOCOX1 Sigma

Cytochrome c Oxidase Assay Kit

sufficient for 100 tests, soluble and membrane bound mitochondria

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Description

Frequently Asked Questions

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Biochem/physiol Actions

Cytochrome c oxidase [EC 1.9.3.1] is located on the inner mitochondrial membrane dividing the mitochondrial matrix from the intermembrane space, and has traditionally been used as a marker for this membrane. It is also located in the cytoplasmic membrane of bacteria. Cytochrome c oxidase provides energy for the cell by coupling electron transport through the cytochrome chain with the process of oxidative phosphorylation.

Features and Benefits

• Simple, optimized protocol - Obtain reproducible results without special training needs
• Useful for determining cytochrome c activity from any mitchondrial source - The enzyme is present in all mitochondria regardless of species
• Useful for detecting the presence of mitochondria in subcellular fraction - Save time and increase confidence in the quality of organelle preparations
• May be used in conjunction with the MITOISO1 Mitochondrial Isolation Kit - Standardized mitochondrial preparation and analysis ensures reproducibility
• Use to analyze the intactness of mitochondrial membranes
• Simple colorimetric measurement of solution

Application

The Cytochrome c Oxidase Assay Kit uses an optimized colorimetric assay based on observation of the decrease in absorbance of ferrocytochrome c measured at 550 nm, which is caused by its oxidation to ferricytochrome c by cytochrome c oxidase.1 This kit is suitable for the detection of mitochondrial outer membrane integrity/mitochondrial stress and for the detection of mitochondria in subcellular fractions.2

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Metabolomic Pathways Chart

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Description

Product #

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Assay Buffer 5× 25 mL MSDS    
Enzyme Dilution Buffer 2× 20 mL MSDS    
Cytochrome c from equine heart, ≥95% (SDS-PAGE) 50 mg MSDS    
Cytochrome c Oxidase positive control 1 vial MSDS    
n-Dodecyl β-D-maltoside, ≥98% (GC) 10 mg MSDS    
DL-Dithiothreitol solution, 1 M .4 mL MSDS    

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1 kit sufficient for 100 reactions (using a 1 ml cuvette), 1 kit sufficient for 480 reactions (using 96 multiwell plates)

Cytochrome c Reductase (NADPH) Assay Kit

1 kit sufficient for 100 tests, determining cytochrome c reductase activity

1 kit sufficient for 50 reactions (in a 2 mL cuvette), 1 kit sufficient for 1,000 reactions (using 96 multiwell plates)

sufficient for 10-20 g (animal tissue), sufficient for 50 assays (2 mL), isolation of enriched mitochondrial fraction from animal tissues

Safety & Documentation

Safety Information

Symbol 
GHS07  GHS07
Signal word 
Warning
Hazard statements 
Precautionary statements 

Protocols & Articles

Articles

Organelle Isolation

The isolation of subcellular fractions by centrifugation is a commonly used technique and is widely applicable across multiple cell and tissue types. Because organelles differ in their size, shape, a...
Mark Frei
BioFiles v6 n5, 22–25
Keywords: Biofiles, Capture ELISA, Cell culture, Centrifugation, Diffusion, Direct immunofluorescence, Dot blot, Enzyme-linked immunosorbent assay, Flow cytometry, Immunoassay, Immunocytochemistry, Immunoelectrophoresis, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Indirect ELISA, Individual protein Immunoprecipitation, Microarray Analysis, Microscopy, Radioimmunoassay, Scanning electron microscopy, Western blot

Peer-Reviewed Papers

References

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1. Fast kinase domain-containing protein 3 is a mitochondrial protein essential for cellular respiration. Simarro M, Gimenez-Cassina A, Kedersha N, et al. Biochem. Biophys. Res. Commun. 401(3), 440-6, (2010)

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2. Identification of poly-ADP-ribosylated mitochondrial proteins after traumatic brain injury. Lai Y, Chen Y, Watkins SC, et al. J. Neurochem. 104(6), 1700-11, (2008)

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Dietary Inorganic Nitrate Improves Mitochondrial Efficiency in Humans. Larsen, F.J., et al. Cell Metab. 13(2), 149-159, (2011)

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Mitochondrial oxygen affinity predicts basal metabolic rate in humans. Larsen, F.J., et al. FASEB J. 25(8), 2843-2852, (2011)

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Galactose Enhances Oxidative Metabolism and Reveals Mitochondrial Dysfunction in Human Primary Muscle Cells. Aguer, C., et al. PLoS ONE 6(12), e28536, (2011)

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Two Sources of Mitochondrial NADPH in the Yeast Saccharomyces cerevisiae. Miyagi, H., et al. J. Biol. Chem. 284(12), 7553-7560, (2009)

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Caveolin-1 Expression Is Essential for Proper Nonshivering Thermogenesis in Brown Adipose Tissue. Cohen, A.W., et al. Diabetes Metab. 54, 679-686, (2005)

Caspase-like proteases involvement in programmed cell death of Phaseolus coccineus suspensor. Lombardi, L., et al. Plant Sci. 172(3), 573-578, (2007)

Transcriptional regulation of pyruvate kinase and phosphoenolpyruvate carboxykinase in the adductor muscle of the oyster Crassostrea gigas during prolonged hypoxia. Le Moullac, G., et al. J. Exp. Zool. 307A(7), 371-382, (2007)

Effect of pesticides on cell survival in liver and brain rat tissues. Astiz, M., et al. Ecotox. Environ. Safety 72(7), 2025-2032, (2009)

Increased susceptibility to oxidative damage in post-diabetic human myotubes. Costford, S.R., et al. Diabetologia 52(11), 2405-2415, (2009)

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Naturally occurring R225W mutation of the gene encoding AMP-activated protein kinase (AMPK)γ3 results in increased oxidative capacity and glucose uptake in human primary myotubes. Crawford, S.A., et al. Diabetologia 53(9), 1986-1997, (2010)

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The Signalling Pathway of CamKII-Mediated Apoptosis and Necrosis in the Ischemia/Reperfusion Injury. Salas, M.A., et al. J. Mol. Cell. Cardiol. 48(6), 1298-1306, (2010)

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Amyloid β-induced Changes in Nitric Oxide Production and Mitochondrial Activity Lead to Apoptosis. Keil, U., et al. J. Biol. Chem. 279(48), 50310-50320, (2004)

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Copper Binding by Tetrathiomolybdate Attenuates Angiogenesis and Tumor Cell Proliferation through the Inhibition of Superoxide Dismutase 1. Juarez, J.C., et al. Clin. Cancer Res. 12, 4974-4982, (2006)

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The Mitochondrial Protease AFG3L2 Is Essential for Axonal Development. Maltecca, F., et al. J. Neurosci. 28(11), 2827-2836, (2008)

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Identification of a New Glycerol-3-phosphate Acyltransferase Isoenzyme, mtGPAT2, in Mitochondria. Lewin, T.M., et al. J. Biol. Chem. 279(14), 13488-13495, (2004)

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Electron transport chain dysfunction in neonatal pressure-overload hypertrophy precedes cardiomyocyte apoptosis independent of oxidative stress. Griffiths, E.R., et al. J. Thorac. Cardiovasc. Surg. 139(6), 1609-1617, (2010)

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Cellular repair of oxidatively induced DNA base lesions is defective in prostate cancer cell lines. Trzeciak, A.R., et al. Carcinogenesis 25(8), 1359-1370, (2004)

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Effects of dietary polyunsaturated fatty acids on mitochondrial metabolism in mammalian hibernation. Gerson, A.R., et al. J. Exp. Biol. 211(16), 2689-2699, (2008)

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Pyrroloquinoline quinone nutritional status alters lysine metabolism and modulates mitochondrial DNA content in the mouse and rat. Bauerly, K.A., et al. Biochim. Biophys. Acta 1760, 1741-1748, (2006)

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Human quadriceps muscle mitochondria: a functional characterization. Rasmussen U.F., and Rasmussen H.N. Mol. Cell Biochem. 208, 37-44, (2000)

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LRP130 Protein Remodels Mitochondria and Stimulates Fatty Acid Oxidation. Liu, L., et al. J. Biol. Chem. 286(48), 41253-41264, (2011)

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Carbon monoxide prevents hepatic mitochondrial membrane permeabilization. Queiroga, C.S.F., et al. BMC Cell Biol. 12, 10, (2011)

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Involvement of SenC in Assembly of Cytochrome c Oxidase in Rhodobacter capsulatus. Swem, D.L., et al. J. Bacteriol. 187(23), 8081-8087, (2005)

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Phosphorylation of Rat Liver Mitochondrial Glycerol-3-phosphate Acyltransferase by Casein Kinase 2. Onorato, T.M., et al. J. Biol. Chem. 280(20), 19527-19534, (2005)

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Detergent-solubilized bovine cytochrome c oxidase: dimerization depends on the amphiphilic environment. Musatov, A. Biochemistry 39, 12996-13004, (2000)

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The Mitochondrial Genome Is a "Genetic Sanctuary" during the Oncogenic Process. Seoane, M., et al. PLoS ONE 6(8), e23327, (2011)

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Increased oxidative stress is associated with balanced increases in hepatocyte apoptosis and proliferation in glycerol-3-phosphate acyltransferase-1 deficient mice. Hammond, L.E., et al. Exp. Mol. Pathol. 82(2), 210-219, (2007)

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Requirements for Nitric Oxide Generation from Isoniazid Activation In Vitro and Inhibition of Mycobacterial Respiration In Vivo. Timmins, G.S., et al. J. Bacteriol. 186(16), 5427-5431, (2004)

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iNOS initiates and sustains metabolic arrest in hypoxic lung adenocarcinoma cells: mechanism of cell survival in solid tumor core. Land, S.C., and Rae, C. Am. J. Physiol. Cell Physiol. 289(4), C918-C933, (2005)

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Phytotoxicity of Sulfamethazine Soil Pollutant to Six Legume Plant Species. Piotrowicz-Cieslak, A. I. J. Toxicol. Environ. Health A 73(17-18), 1220-1229, (2010)

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Effects of Low-Level Light Therapy on Hepatic Antioxidant Defense in Acute and Chronic Diabetic Rats. Lim, J., et al. J. Biochem. Mol. Toxicol. 23(1), 1-8, (2009)

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Preconditioning tachycardia decreases the activity of the mitochondrial permeability transition pore in the dog heart. Sánchez, G., et al. Biochem. Biophys. Res. Commun. 410(4), 916-921, (2011)

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