5 5 dithiobis 2 nitrobenzoic acid

Manufactured by Sisco Research Laboratories

Sourced in India

5,5-dithiobis(2-nitrobenzoic acid) is a chemical compound used as a reagent in various laboratory applications. It functions as a sulfhydryl (thiol) group detecting agent.

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14 protocols using 5 5 dithiobis 2 nitrobenzoic acid

Antioxidant Assay Protocol

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Nitro blue tetrazolium (NBT), 2,4-dinitrophenyl hydrazine (DNPH), 5,5′-dithio-bis-(2-nitrobenzoic acid) (DTNB), dithiothreitol (DTT), 1-diphenyl-2-picrylhydrazyl (DPPH) radical, polyvinyl pyrrolidone, nicotinamide adenine dinucleotide phosphate reduced (NADPH), bovine serum albumin, and methionine were provided by Sisco Research Laboratories Pvt., Ltd. (Mumbai, India). Thiobarbituric acid (TBA) was purchased from Molychem (Mumbai, India). Antioxidant assay kit and Sephadex G-25 column were purchased from Cayman Chemical Company (MI, USA) and Sigma-Aldrich (MO, USA), respectively. Ethanol and α-tocopherol were obtained from Changshu Yangyuan Chemical (Jiangsu, China) and Acros Organics (NJ, USA), respectively. All the other chemicals employed were of standard analytical grade and provided by Thermo Fisher Scientific India Pvt., Ltd. (Mumbai, India).

Prathyusha I.V.S.N, & Chaitanya K.V. (2019). Effect of water stress on the physiological and biochemical responses of two different Coleus (Plectranthus) species. Biologia futura, 70(4).

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Biochemical Assay Protocol

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Pyrithiamine hydrobromide was procured from Sigma (MO, USA). 5, 5’-dithiobis-2-nitrobenzoic acid, (DTNB), ethylene diamine tetraacetic acid (EDTA), tris hydrochloride, trichloroacetic acid (TCA), 2-thiobarbituric acid (TBA) were purchased from Sisco research laboratories (Mumbai, India). All other chemicals were used of analytical grade.

Sharma A, & Bist R. (2019). Alteration in MDA, GSH level and hematological changes due to thiamine deficiency in Mus musculus. Interdisciplinary Toxicology, 11(4), 321-325.

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Antioxidant and Oxidative Stress Assays

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ABTS [2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)], 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) was purchased from Roche diagnostics (Mannheim, Germany) and Fluka (Buchs, Switzerland), respectively. Nitro blue tetrazolium (NBT), 2-deoxy-2-ribose, sodium nitroprusside (SNP), phenazine methosulfate (PMS), 1,10-phenanthroline, 5,5′-dithiobis-2-nitrobenzoic acid (DTNB), lipoic acid, ferrozine and bathophenanthrolinesulfonate disodium salt were purchased from Sisco Research Laboratories Pvt. Ltd, Mumbai, India. Folin-ciocalteu reagent, 2,4-dinitrophenylhydrazine (DNPH), butylated hydroxyltoluene (BHT), 1-chloro-2,4-dinitrobenzene (CDNB), chloramine-T, Dimethyl-4-aminobenzaldehyde and hydroxylamine hydrochloride were taken from Merck, Mumbai, India. Spectrochem Pvt. Ltd, (India) supplied Diethylenetriaminepentaacetic acid (DTPA). Iron-dextran and guanidine hydrochloride was obtained from Sigma-Aldrich, USA. Desirox (Deferasirox) was purchased from Cipla Ltd., India. All the other used reagents are molecular biology grade and obtained from reputed suppliers.

Chaudhuri D., Ghate N.B., Panja S., Das A, & Mandal N. (2015). Wild Edible Fruit of Prunus nepalensis Ser. (Steud), a Potential Source of Antioxidants, Ameliorates Iron Overload-Induced Hepatotoxicity and Liver Fibrosis in Mice. PLoS ONE, 10(12), e0144280.

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Biochemical Assay Protocols for Research

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The 1-chloro-2,4-dinitrobenzene, ammonium sulphate, 2,4-dinitrophenylhydrazine, ethylene diaminetetra acetic acid, 5,5-dithiobis(2-nitrobenzoic acid), N-ethylmaleimide, nitroblue tetrazolium, reduced nicotinamide adenine dinucleotide, potassium dihydrogen phosphate, phenazinemethosulphate, sodium pyrophosphate, reduced glutathione, sodium azide, thiobarbituric acid, 5-thio-2-nitrobenzoic acid and trichloro acetic acid were purchased from Sisco Research Laboratory, Mumbai, India. Bradford reagent, bovine serum albumin and cadmium chloride were procured from Sigma-Aldrich Chemical Company, St. Louis, USA. All antibodies for immunoblotting were purchased from Sigma-Aldrich Chemical Company St. Louis, USA. HPLC grade solvents (acetic acid, formic acid, methanol and acetonitrile) were procured from Merck, Mumbai, India.

Dua T.K., Dewanjee S., Khanra R., Bhattacharya N., Bhaskar B., Zia-Ul-Haq M, & De Feo V. (2015). The effects of two common edible herbs, Ipomoea aquatica and Enhydra fluctuans, on cadmium-induced pathophysiology: a focus on oxidative defence and anti-apoptotic mechanism. Journal of Translational Medicine, 13, 245.

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Antioxidant Activity Evaluation

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Bovine serum albumin and bleomycin were obtained from Sigma-Aldrich (St. Louis, MO, USA). Trichloroacetic acid was acquired from Merck (Mumbai, India), and thiobarbituric acid (TBA) and nitroblue tetrazolium chloride were purchased from Loba Chemie Pvt. Ltd., (Mumbai, India). 5,5′-Dithiobis(−2-nitro benzoic acid), phenazonium methosulfate, nicotinamide adenine dinucleotide and reduced glutathione (GSH) were purchased from Sisco Research Laboratories Pvt., Ltd., (Mumbai, India). All other chemicals and reagents used were of the highest analytical grade.

ALAM B., MAJUMDER R., AKTER S, & LEE S.H. (2014). Piper betle extracts exhibit antitumor activity by augmenting antioxidant potential. Oncology Letters, 9(2), 863-868.

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Protein Quantification and Modification Analysis

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Fatty acid and globulin-free
HSA and 2,4,6-trinitrobenzene sulfonic acid (TNBSA) were obtained
from Sigma-Aldrich, USA. Thiamine and 5,5-dithiobis(2-nitrobenzoic
acid) were purchased from Sisco Research Laboratories, India. 2,4-dinitrophenylhydrazine
was obtained from HiMedia Laboratories (India). All the other chemicals
used were of analytical grade.

Abdullah K.M., Arefeen A., Shamsi A., Alhumaydhi F.A, & Naseem I. (2021). Insight into the In Vitro Antiglycation and In Vivo Antidiabetic Effects of Thiamine: Implications of Vitamin B1 in Controlling Diabetes. ACS Omega, 6(19), 12605-12614.

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Geraniin Antioxidant and Cytotoxic Assays

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Dulbecco’s modified essential media (DMEM), Foetal Bovine Serum (FBS), trypsin and Penicillin-streptomycin were purchased from Gibco. TBH, 2-thiobarbituric acid (TBA), 2′,7′-dichlorofluorescein diacetate (DCFH-DA) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma Aldrich, USA. Reduced glutathione, potassium phosphate, sodium carbonate, trichloroacetic Acid (TCA) and 5,5′-dithio-bis (2 nitrobenzoic acid) (DTNB) were obtained from Sisco Research Laboratory, India. Other common chemicals used in our studies were of the highest quality, commercially available from local suppliers. Geraniin (purity of > 90%) was procured from AJ organic Pvt. Ltd., Pune, India.

Aayadi H., Mittal S.P., Deshpande A., Gore M, & Ghaskadbi S.S. (2017). Cytoprotective effect exerted by geraniin in HepG2 cells is through microRNA mediated regulation of BACH-1 and HO-1. BMB Reports, 50(11), 560-565.

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Biochemical Assays for Cellular Experiments

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Bovine serum albumin (BSA), Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), Bradford reagent, Collagenase type I, antibodies for immunoblotting were purchased from Sigma-Aldrich Chemical Company (St. Louis, MO), USA. Solvents (HPLC grade) were obtained from Merck, Mumbai, India. Kits for measurement of serum biochemical parameters were purchased from Span diagnostic Ltd., India. 1-Chloro-2,4-dinitrobenzene, ammonium sulphate, sodium arsenite (NaAsO₂), 2,4-dinitrophenylhydrazine, ethylene diaminetetraacetic acid, 5,5-dithiobis(2-nitrobenzoic acid), N-ethylmaleimide, nitro blue tetrazolium, reduced nicotinamide adenine dinucleotide, potassium dihydrogen phosphate, phenazinemethosulphate, sodium pyrophosphate, reduced glutathione, sodium azide, thiobarbituric acid, 5-thio-2-nitrobenzoic acid and trichloro acetic acid were purchased from Sisco Research Laboratory, Mumbai, India.

Dua T.K., Dewanjee S., Gangopadhyay M., Khanra R., Zia-Ul-Haq M, & De Feo V. (2015). Ameliorative effect of water spinach, Ipomea aquatica (Convolvulaceae), against experimentally induced arsenic toxicity. Journal of Translational Medicine, 13, 81.

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Spectrophotometric Assay for Antioxidant Enzymes

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Bathophenanthroline sulfonate disodium salt, 5,5′-dithiobis-2-nitrobenzoic acid (DTNB) and N-(1-Naphthyl) ethylenediamine dihydrochloride (NED) were procured from Sisco Research Laboratories Pvt. Ltd, India. 1-chloro-2,4-dinitrobenzene (CDNB), Dimethyl-4-aminobenzaldehyde, and N,N- dimethyl-4-nitrosoaniline ammonium iron (II) sulfate hexahydrate ((NH₄)₂Fe(SO₄)_2, 6H₂O) were obtained from Merck, Mumbai, India. Guanidine hydrochloride and Iron-dextran was purchased from Sigma-Aldrich, USA. Cipla Ltd., India provided Desirox (Generic name-Deferasirox). All of the other used reagents were of molecular biology grade and were obtained from reputable suppliers.

Chaudhuri D., Ghate N.B., Panja S, & Mandal N. (2016). Role of phenolics from Spondias pinnata bark in amelioration of iron overload induced hepatic damage in Swiss albino mice. BMC Pharmacology & Toxicology, 17, 34.

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Cadmium Chloride Induced Cellular Stress Response

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CA, cadmium chloride (CdCl₂), fetal bovine serum (FBS), Bradford reagent, bovine serum albumin (BSA), and cell culture media were obtained from Sigma-Aldrich Chemical Company, St. Louis, USA. Antibodies for Western blotting were obtained from Cell Signaling Technology, Beverley, USA, Denver, USA, and Bio-Rad, Hercules, USA. (NH₄)₂SO₄, 2,4-dinitrophenylhydrazine, CCl₃COOH, 5-thio-2-nitrobenzoic acid, 5,5-dithiobis(2-nitrobenzoic acid), 1-chloro-2,4-dinitrobenzene, ethylene diaminetetraacetic acid (EDTA), Na₄P₂O₇, NaN₃, KH₂PO₄, N-ethylmaleimide, nitro blue tetrazolium (NBT), phenazinemethosulphate, reduced glutathione (GSH), reduced nicotinamide adenine dinucleotide (NADH), and thiobarbituric acid (TBA) were bought from Sisco Research Laboratory, Mumbai, India. Solvents were bought from Merck, Mumbai, India. Kits for measurement of different biochemical parameters were purchased from Span diagnostic Ltd., Kolkata, India. ELISA kits were obtained from R&D Systems, Minneapolis, USA and Thermo Fisher Scientific Co., Waltham, USA.

Das S., Dewanjee S., Dua T.K., Joardar S., Chakraborty P., Bhowmick S., Saha A., Bhattacharjee S, & De Feo V. (2019). Carnosic Acid Attenuates Cadmium Induced Nephrotoxicity by Inhibiting Oxidative Stress, Promoting Nrf2/HO-1 Signalling and Impairing TGF-β1/Smad/Collagen IV Signalling. Molecules, 24(22), 4176.

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