Dithionitrobenzoic acid

Manufactured by Merck Group

Sourced in United States

Dithionitrobenzoic acid is a chemical compound commonly used in biochemistry and molecular biology laboratories. It serves as a reagent for the quantitative determination of free sulfhydryl groups in proteins and peptides.

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Dithionitrobenzoic acid (DTNB), (2 citations)

4 protocols using dithionitrobenzoic acid

Oxidative Stress Enzyme Assay

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Benznidazole (BZL), dithionitrobenzoic acid, oxidized glutathione (GSSH), reduced glutathione (GSH), glutathione, nitroblue tetrazolium, MK571, β-NADPH, riboflavin, rifampicin (RIF), sulfosalicylic acid, tert-butyl hydroperoxide (tBOOH) and 2-vinylpyridine were purchased from Sigma-Aldrich (St. Louis, MO, USA). DMSO and hydrogen peroxide were purchased from Merck (Darmstadt, HE, Germany). All other chemicals were of analytical grade purity.

Rigalli J.P., Perdomo V.G., Ciriaci N., Francés D.E., Ronco M.T., Bataille A.M., Ghanem C.I., Ruiz M.L., Manautou J.E, & Catania V.A. (2016). The antitripanocide Benznidazole promotes adaptive response to oxidative injury: Involvement of the nuclear factor-erythroid 2-related factor-2 (Nrf2) and multidrug resistance associated protein 2 (MRP2). Toxicology and applied pharmacology, 304, 90-98.

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Purification and Characterization of Recombinant Proteins

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AdoCbl, ATP, GTP, PPPi, and dithionitrobenzoic acid were from Sigma-Aldrich. The primers used for mutagenesis were purchased from Integrated DNA Technologies. IPTG and Tris (2-carboxyethyl) phosphine (TCEP) were from gold biotechnology. Nickel-nitrilotriacetic acid (Ni(II)-NTA) resin was from Qiagen.

Gouda H., Mascarenhas R., Pillay S., Ruetz M., Koutmos M, & Banerjee R. (2021). Patient mutations in human ATP:cob(I)alamin adenosyltransferase differentially affect its catalytic versus chaperone functions. The Journal of Biological Chemistry, 297(6), 101373.

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Synthetic Chemistry Reagent Usage

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All chemical reagents involved in synthetic procedures were purchased commercially from J&K Chemicals. The chemical reagents used in activity measurement, including dimethyl sulfoxide (DMSO), ethylene diamine tetraacetic acid (EDTA), dithionitrobenzoic acid (DTNB), and MTT, were purchased from Sigma-Aldrich Inc.

Li G., Guo Q., Feng C., Chen H., Zhao W., Li S., Hong Y, & Sun D. (2021). Synthesis of oxadiazole-2-oxide derivatives as potential drug candidates for schistosomiasis targeting SjTGR. Parasites & Vectors, 14, 225.

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Glutathione Redox State Evaluation

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GSSG/GSH was assessed by mixing in a cuvette 0.05 M Na-phosphate buffer (pH 7.0), 1 mM EDTA (pH 7.0) and 10 mM dithionitrobenzoic acid (Sigma-Aldrich) plus an aliquot of the sample. After 2 min of reaction, 24:6 GSH content was evaluated by reading absorbance at 412 nm, calculated referring to a standard curve and expressed as µg/mg protein. Suitable volumes of diluted glutathione reductase and of reduced nicotinamide adenine dinucleotide phosphate were then added to convert the oxidized glutathione to the reduced form and then evaluate the total glutathione level. The difference between total glutathione and GSH content represents the GSSG content, also expressed as µg/mg protein.

Marano F., Frairia R., Rinella L., Argenziano M., Bussolati B., Grange C., Mastrocola R., Castellano I., Berta L., Cavalli R, & Catalano M.G. (2017). Combining doxorubicin-nanobubbles and shockwaves for anaplastic thyroid cancer treatment: preclinical study in a xenograft mouse model. Endocrine-related cancer, 24(6).

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