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Xanthine oxidase from bovine milk

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Xanthine oxidase from bovine milk is an enzyme that catalyzes the oxidation of hypoxanthine to xanthine and then to uric acid. It is a molybdoflavoprotein that contains flavin adenine dinucleotide (FAD) and molybdenum as cofactors. Xanthine oxidase plays a role in purine metabolism.

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Lab products found in correlation

Xanthine, by Merck Group (9 mentions) Allopurinol, by Merck Group (3 mentions) 2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (2 mentions) Thiobarbituric acid, by Merck Group (2 mentions) Adenosine, by Merck Group (2 mentions) Inosine, by Merck Group (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Butylated hydroxy toluene, by Thermo Fisher Scientific (2 mentions) Absolute ethanol, by ITW Reagents (2 mentions) Hydrochloric acid (hcl), by Merck Group (2 mentions) Potassium cyanide, by Merck Group (2 mentions) 2 thiobarbituric acid, by Thermo Fisher Scientific (2 mentions) Ethylenediaminetetraacetic acid disodium salt edta 2na, by GE Healthcare (2 mentions) Formic acid, by Thermo Fisher Scientific (1 mentions) Trifluoroacetic acid, by Thermo Fisher Scientific (1 mentions) Aniline, by Merck Group (1 mentions) Sodium dithionite, by Merck Group (1 mentions) Nabh3cn, by Merck Group (1 mentions) Methyl viologen dichloride, by Merck Group (1 mentions) 2 deoxy d ribose, by Merck Group (1 mentions)

Close spelling variants of this product

Xanthine (244 citations) Xanthine oxidase (219 citations) Bovine milk (19 citations) Xanthine oxidase from (4 citations) Bovine milk xanthine oxidase (2 citations) Bovine xanthine oxidase (2 citations) Xanthine oxidase (XOD) from bovine milk (2 citations)

21 protocols using xanthine oxidase from bovine milk

1

Synthesis and Purification of Iodotyrosine Derivatives

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3-Fluoro-l-tyrosine (F-Tyr) was obtained from AstaTech, Inc (Bristol, PA). 3-Iodo-l-tyrosine (I-Tyr), O-methyl tyrosine, formic acid (88%) and trifluoroacetic acid were obtained from Acros Organics (Morris Plains, NJ). 3,4-Dimethylaniline, aniline, 2′-deoxy-d-ribose, NaBH3CN, xanthine oxidase from bovine milk, methyl viologen dichloride and sodium dithionite were obtained from Sigma-Aldrich (Madison, WI). SilicaFlash P60 silica (SiliCycle Inc., Quebec City, Quebec) was used for all column chromatography. Human IYD (HsIYD) lacking its N-terminal transmembrane sequence (residues 2–31) was expressed as a SUMO fusion, purified by Ni2+ affinity and size exclusion chromatography as described previously.16 (link) IYD from Thermotoga neapolitana (TnIYD) was expressed with a C-terminal His6-tag and purified by Ni2+ affinity chromatography as described previously.17 (link) Syntheses of 2′-deoxyFMN, O-Me I-Tyr and I-aminoPhe are described in the ESI.†
Kozyryev A., Boucher P.A., Quiñones-Jurgensen C.M, & Rokita S.E. (2023). The 2′-hydroxy group of flavin mononucleotide influences the catalytic function and promiscuity of the flavoprotein iodotyrosine dehalogenase. RSC Chemical Biology, 4(9), 698-705.
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2

Inflammatory Signaling Pathway Assays

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All the cell culture reagents were obtained from HyClone Laboratories (South Logan, UT, USA). Xanthine oxidase from bovine milk, xanthine substrate, indomethacin, N-acetylcysteine, prednisolone, 4-Nitro blue tetrazolium chloride (NBT), 2,2-Diphenyl-1-picrylhydrazyl (DPPH), carrageenan and lipopolysaccharides (LPS) were procured from Sigma–Aldrich (St Louis, MO, USA). Diphenyleneiodonium (DPI) chloride, lucigenin, and NADPH tetrasodium was purchased from Enzo life sciences (Farmingdale, NY, USA). The primary antibodies against total and phosphorylated forms of ERK1/2, p38 MAPK and JNK1/2, and the secondary antibodies conjugated with horseradish peroxidase (HRP) were acquired from Cell Signaling Technology Inc. (Beverly, MA, USA). Anti-β-actin and anti-COX-2 were provided by BD Biosciences (San Jose, CA, USA) and Sigma–Aldrich, respectively. The anti-rabbit and anti-mouse secondary antibodies conjugated with horseradish peroxidase (HRP) were acquired from Cell Signaling Technology Inc (Beverly, MA, USA).
Nguyen T.D., Nguyen T.H., Do T.H., Tran V.T., Nguyen H.A, & Pham D.V. (2022). Anti-inflammatory effect of a triterpenoid from Balanophora laxiflora: results of bioactivity-guided isolation. Heliyon, 8(3), e09070.
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3

Antioxidant and Hepatoprotective Evaluation

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Ethanol, nitroblue tetrazolium salt, xanthine, copper chloride, glutathione, xanthine oxidase from bovine milk (0.1–0.4 units/mg of protein), glutathione reductase from baker’s yeast (Saccharomyces cerevisiae, 100–300 units/mg of protein), and silymarin (SIL) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA). F-12 medium, fetal bovine serum (FBS) and penicillin/streptomycin were obtained from Hyclone (Logan, UT, USA). Antibodies against ADH, ALDH1/2, CYP2E1, SOD, GPx, β-actin, and horseradish peroxidase-conjugated anti-goat IgG were purchased from Santa Cruz Biotech (Santa Cruz, CA, USA). Anti-phospho-JNK, JNK, phospho-ERK, ERK, phospho-p38, p38, and horseradish peroxidase-conjugated anti-rabbit IgG were purchased from Cell Signaling technology Inc. (Beverly, MA, USA). CAT antibody was purchased from Abcam (Cambridge, MA, USA). All other chemicals in this study were analytical grade.
Bak M.J., Truong V.L., Ko S.Y., Nguyen X.N., Ingkasupart P., Jun M., Shin J.Y, & Jeong W.S. (2016). Antioxidant and Hepatoprotective Effects of Procyanidins from Wild Grape (Vitis amurensis) Seeds in Ethanol-Induced Cells and Rats. International Journal of Molecular Sciences, 17(5), 758.
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4

Graphene Oxide Synthesis and Characterization

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GO was prepared using a modified Hummers method and purified as described previously.68 RGO was produced by heating multilayer GO flakes at 250 C for 30 min in nitrogen flow. Few layer graphene (3–5 layers with nominal lateral dimension of 800 nm) was obtained commercially and characterized. Ascorbic acid, titanium (IV) oxide (TiO2) nanopowders (<25 nm particle size, >99.5 % trace metal basis), iron (II) sulfate haptahydrate (FeSO4·7H2O), reduced GSH, D-mannitol, pC60, xanthine, xanthine oxidase from bovine milk grade IV,2,2′-azobis(2-amidinopropane) hydrochloride (AAPH), thiobarbituric acid (TBA), trichloricacetic acid (TCA), DPPH, 2-deoxy-D-ribose (deoxyribose), sodium dodecyl sulfate (SDS), ABTS, β-Nicotinamide adenine dinucleotide 23-phosphate reduced tetrasodium salt hydrate (NADH), phenazinemethosulfate (PMS), xanthine, xanthine oxidase from bovine milk, and NBT were purchased from Sigma Chemicals Co (St. Louis, MO). Hydrogen peroxide (H2O2), phosphate buffered saline (PBS) (pH 7.4), and EDTA were purchased from Fisher Scientific Inc. (Pittsburgh, PA). 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) were purchased from Dojindo Molecular Technologies, Inc. (Rockville, MD). ThioGlo-1 Fluorescent Thiol Reagent is purchased from EMD Millipore Chemical (Darmstadt, Germany), Nanopure water was used throughout.
Qiu Y., Wang Z., Owens A.C., Kulaots I., Chen Y., Kane A.B, & Hurt R.H. (2014). Antioxidant Chemistry of Graphene-Based Materials and its Role in Oxidation Protection Technology. Nanoscale, 6(20), 11744-11755.
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5

Serotonin Biotin Conjugate Synthesis

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Serotonin and dithiothreitol (DTT) were purchased from Wako Pure Chemicals. Human myeloperoxidase was obtained from Planta Natural Products. Xanthine oxidase from bovine milk (XOD; type X4500) and GAPDH (from rabbit) were purchased from Sigma. Acetaldehyde was purchased from Merck. CanGetSignal-1 and -2 were purchased from TOYOBO. Tris[2-carboxyethyl] phosphine hydrochloride (TCEP) was purchased from Nacalai Tesque Inc. Biotinylated Serotonin (Serotonin–biotin) was prepared by reacting sulfo-NHS-LC-biotin (Thermo Scientific) with Serotonin [13] (link).
Kato Y., Ono S., Kitamoto N, & Kettle A.J. (2014). Covalent modification of cytoskeletal proteins in neuronal cells by tryptamine-4,5-dione. Redox Biology, 2, 983-990.
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6

Synthesis and Purification of m7Guo

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Formycin A was from Berry & Associates, Inc. (USA), adenosine, inosine, guanosine and xanthine oxidase from bovine milk (1 U/mg) were obtained from Sigma-Aldrich (USA). 7-methylguanosine (m7Guo) was synthesised from guanosine according to Jones’ and Robins' method involving methyl iodide25 (link). This yields the preparation free from sulfate, which as an ion resembling phosphate could bias the results. All other chemicals were from Roth (Germany) or Sigma-Aldrich (USA), and were of the highest available purity. Background contaminant phosphate in buffers and other chemicals used in this study was measured spectrophotometrically using the method involving phosphomolybdate complex reduced by ascorbic acid26 (link).
Narczyk M., Mioduszewski Ł., Oksiejuk A., Winiewska-Szajewska M., Wielgus-Kutrowska B., Gojdź A., Cieśla J, & Bzowska A. (2021). Single tryptophan Y160W mutant of homooligomeric E. coli purine nucleoside phosphorylase implies that dimers forming the hexamer are functionally not equivalent. Scientific Reports, 11, 11144.
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7

Antioxidant Activity Assay Protocol

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xanthine oxidase from bovine milk, luminol sodium salt, xanthine, oxypurinol, PBS tabs, Na-EDTA salt, gelatin from bovine skin, penicillin/streptomycin, trypsin-EDTA, Trolox, and 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sodium perborate, boric acid, NaOH, and FeCl2 were from Carlo Erba (Milan, Italy). M200 medium, low serum growth supplements, and fetal bovine serum, RNaseOUT, were purchased from Thermo Fisher Scientific (Waltham, MA, USA). RNeasy Mini Kit was from QIAGEN (Hilden, Germany). Primers for RT-PCR were purchased from IDT (Coralville, IA, USA). Cell counting kit-8 (CCK8) and LDH assay kit were purchased from Dojindo Molecular Technologies (Rockville, MD, USA). SuperScript® III First-Strand Synthesis SuperMix and EXPRESS SYBR® GreenER™ qPCR SuperMix were purchased from Life Technologies (Carlsbad, CA, USA). All the other chemicals and solvents were of the highest analytical grade.
Caliceti C., Capriotti A.L., Calabria D., Bonvicini F., Zenezini Chiozzi R., Montone C.M., Piovesana S., Zangheri M., Mirasoli M., Simoni P., Laganà A, & Roda A. (2019). Peptides from Cauliflower By-Products, Obtained by an Efficient, Ecosustainable, and Semi-Industrial Method, Exert Protective Effects on Endothelial Function. Oxidative Medicine and Cellular Longevity, 2019, 1046504.
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8

Purification and Synthesis of Biochemical Reagents

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Wild-type PNP was obtained and purified as previously described [34 (link)]. Adenosine, inosine, and xanthine oxidase from bovine milk (1 U/mg) were obtained from Sigma-Aldrich (St. Louis, MO, USA). The 7-methylguanosine was synthesized from guanosine according to Jones’ and Robins’ method involving methyl iodide [35 (link)]. This approach assured the preparation was free from sulphate, which as an ion-resembling phosphate could bias the results. All other chemicals were purchased from Roth (Karlsruhe, Germany) or Sigma-Aldrich and were of the highest available purity.
Dyzma A., Wielgus-Kutrowska B., Girstun A., Matošević Z.J., Staroń K., Bertoša B., Trylska J, & Bzowska A. (2023). Trimeric Architecture Ensures the Stability and Biological Activity of the Calf Purine Nucleoside Phosphorylase: In Silico and In Vitro Studies of Monomeric and Trimeric Forms of the Enzyme. International Journal of Molecular Sciences, 24(3), 2157.
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9

Xanthine Oxidase Inhibition by E. characias

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The inhibitory effect of E. characias extracts on xanthine oxidase activity was determined spectrophotometrically by monitoring the formation of uric acid at 295 nm. The reaction mixture contained 879 μL of 100 mM phosphate buffer, pH 7.5, 50 μL of an aqueous solution of xanthine oxidase from bovine milk (0.5 U/mL, Sigma Chemical Co.), and 10 μL of extract sample solution or control sample solution (ethanol or water). After mixing, 61 μL of xanthine solution 0.82 mM was added and the enzyme activity was determined at 295 nm for 3 min at 25°C. Allopurinol was used as positive control.
Fais A., Era B., Di Petrillo A., Floris S., Piano D., Montoro P., Tuberoso C.I., Medda R, & Pintus F. (2018). Selected Enzyme Inhibitory Effects of Euphorbia characias Extracts. BioMed Research International, 2018, 1219367.
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10

Characterization of Bioactive Compounds

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Chlorogenic acid, coumarin, 3,4-dicaffeoylquinic acid, cinnamaldehyde, trans-cinnamic acid, and o-methoxycinnamaldehyde were purchased from ChemFaces (China), respectively. The purities of them were determined to be 98% by high performance liquid chromatography (HPLC) analysis. Acetonitrile, methanol, and water were purchased from J. T. Baker (Phillipsburg, NJ, USA). Trifluoroacetic acid, allopurinol, dimethyl sulfoxide, xanthine oxidase from bovine milk (0.09 units/mg solid), xanthine sodium salt, and phosphate buffer (pH 7.2) were obtained from Sigma-Aldrich (St. Louis, MO, USA).
Lee Y.S., Son E., Kim S.H., Lee Y.M., Kim O.S, & Kim D.S. (2017). Synergistic Uric Acid-Lowering Effects of the Combination of Chrysanthemum indicum Linne Flower and Cinnamomum cassia (L.) J. Persl Bark Extracts. Evidence-based Complementary and Alternative Medicine : eCAM, 2017, 9764843.
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