Indomethacin capsules (Munawar Pharma PVT Ltd), diclofenac sodium tablets (Novartis Pharma PAK Ltd), carrageenan (M/s Bio Basic, Canada), Freund's Complete Adjuvant (M/s Sigma Aldrich, St. Louis USA). 1-Chloro-2,4-dinitrobenzene (CDNB), pyrogallol, sodium chloride, and tetramethylpropane (TMP) were supplied by Sigma Aldrich Co. Ltd, Korea. Di-sodium hydrogen phosphate, hydrochloric acid, potassium dichromate, and sodium potassium tartrate were supplied by Reidel_deHaien, Sigma Aldrich. Copper sulfate pentahydrate, hydrogen peroxide, potassium dihydrogen phosphate, and reduced glutathione (GSH) were supplied from Merck, Germany. For alkaline phosphatase (AP), alanine transaminase (ALT), and aspartate transaminase (AST) analysis kits by Crescent Diagnostics were used. For TNF-α analysis, a kit by Glory Science Co., Ltd was used.
Reduced glutathione gsh
Manufactured by Merck Group
Sourced in United States, Germany, India, China
Reduced glutathione (GSH) is a tripeptide composed of glutamic acid, cysteine, and glycine. It is a common antioxidant found in living organisms, playing a crucial role in cellular processes. GSH functions as a reducing agent, helping to maintain the appropriate oxidation-reduction state within cells.
Automatically generated - may contain errors
Lab products found in correlation
Oxidized glutathione gssg, by Merck Group (6 mentions)
Thiobarbituric acid, by Merck Group (6 mentions)
Glutathione reductase, by Merck Group (5 mentions)
Trichloroacetic acid, by Merck Group (5 mentions)
Sodium azide, by Merck Group (4 mentions)
Thiobarbituric acid tba, by Merck Group (4 mentions)
5 5 dithiobis 2 nitrobenzoic acid dtnb, by Merck Group (4 mentions)
Quercetin, by Merck Group (4 mentions)
1 chloro 2 4 dinitrobenzene cdnb, by Merck Group (4 mentions)
Disodium hydrogen phosphate, by Merck Group (3 mentions)
5 5 dithiobis 2 nitrobenzoic acid, by Merck Group (3 mentions)
1 chloro 2 4 dinitrobenzene, by Merck Group (3 mentions)
Hydrogen peroxide h2o2, by Merck Group (3 mentions)
Bovine serum albumin (bsa), by Merck Group (3 mentions)
Ethylene diamine tetraacetic acid (edta), by Merck Group (2 mentions)
Nadph, by Merck Group (2 mentions)
Methionine, by Merck Group (2 mentions)
N acetylcysteine, by Merck Group (2 mentions)
L cysteine, by Merck Group (2 mentions)
Tris hcl, by Merck Group (2 mentions)
73 protocols using reduced glutathione gsh
1
Anti-inflammatory Compound Evaluation Protocol
2
Glutathione Peroxidase Activity Measurement
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The glutathione peroxidase (Gpx) activity was assayed in plasma, according to Lawrence and Burk [46 (link)] and liver and dorsal muscle as described by Fontagné-Dicharry et al. [21 (link)]. Gpx activity in plasma samples was evaluated immediately after thawing. In the case of liver and muscle, samples were rapidly thawed and homogenized in 10 volumes (w/v) of ice-cold saline for 3 min and centrifuged for 15 min at 4000× g and the supernatants collected to evaluate the activity of GPx. Gpx activity present in the supernatants was measured in a solution of 50 mM phosphate buffer (pH 7.4), 1 mM EDTA (Merck, Darmstadt, Germany), 2 mM sodium azide (Merck, Darmstadt, Germany), 2 mM reduced glutathione (GSH) (Merck, Darmstadt, Germany), 0.1 mM NADPH (Merck, Darmstadt, Germany), and 0.2 mM glutathione reductase (Merck, Darmstadt, Germany) following the reduction of H2O2 (50μM) at 30 °C and 340 nm. One unit of Gpx activity was reported as l mol NADPH consumed per min per mg of plasma protein, using the appropriate molar absorptivity coefficient for NADPH (6220 mol L−1 cm−1). Plasma proteins were measured by the method of Lowry et al. [47 (link)].
3
Ursolic Acid Bioactivity Evaluation
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Ursolic acid (purity ≥ 98%), thiobarbituric acid (TBA), phenazinemethosulphate (PMS), nitroblue tetrazolium (NBT), 5,5-dithiobis 2-nitrobenzoic acid (DTNB), 3-(4, 5-dimethyl-2-thiaozolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT), 2′7′-diacetyl dichlorofluorescein (DCFH-DA), rhodamine-123, and nicotinamide adenine dinucleotide (NAD) were supplied by Sigma-Aldrich, St. Louis, USA. The mouse monoclonal antibodies anti-TNF-α, anti-NF-κB (p65), anti-p53, anti-Bax, anti-Bcl-2, anti-caspase-3, anti-MMP-2, anti-MMP-9, anti-β-actin, and goat anti-mouse IgG-HRP polyclonal antibody were also purchased from Sigma-Aldrich, St. Louis, USA. RNA isolation kit, cDNA synthesis kit, PCR primers, and PCR master mix were purchased from Qiagen, USA. Bovine serum albumin (BSA), RIPA buffer, low melting agarose (LMPA), normal melting agarose (NMPA), phosphate buffered saline (PBS) and reduced glutathione (GSH) were purchased from Merck, India. All other chemicals, gradient solvents, and other analytical grades were acquired from S.D Fine Chemical and Himedia, India.
4
Measuring Glutathione Peroxidase Activity
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The Gpx activity was measured in plasma following the indications of Lawrence and Burk [79 (link)] and in the liver and in the dorsal muscle those of Fontagné-Dicharry et al. [80 (link)]. For plasma, Gpx activity was evaluated immediately after thawing. For liver and muscle, after thawing the samples on ice and homogenizing them in 10 volumes (w/v) of ice-cold saline for 3 min, they were centrifuged for 15 min at 4000× g before the activity of GPx was determined in the supernatants. GPx activity was measured in a solution of 50 mM phosphate buffer (pH 7.4), 1 mM EDTA (Merck, Darmstadt, Germany), 2 mM sodium azide (Merck, Darmstadt, Germany), 2 mM reduced glutathione (GSH) (Merck, Darmstadt, Germany), 0.1 mM NADPH (Merck, Darmstadt, Germany), and 0.2 mM glutathione reductase (Merck, Darmstadt, Germany). H2O2 (50 μM) reduction at 30 °C was measured at 340 nm in an Epoch spectrophotometer. One unit of Gpx activity was valued as 1 mol NADPH consumed per min per mg of plasmatic proteins, using the appropriate molar absorptivity coefficient for NADPH (6220 mol L−1 cm−1). Plasmatic protein measurement was performed following the method of Lowry et al. [81 (link)].
5
Fab and F(ab')2 Fragment Preparation
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Fab and F(ab’)2 fragments were generated using the Pierce Fab preparation kit (Thermo Fisher) or FragIT kit (Genovis, Lund, Sweden) according to the manufacturer's protocols. Fab and F(ab’)2 fragments were dialyzed to artificial CSF, concentrated at 10 mg/mL, and filter-sterilized. FAE was performed as previously described.19 (link),23 (link) IgG4 or IgG1 from Nfasc155-reactive patients were incubated with a 3-fold excess of healthy control IgG4 in the presence or absence of 0.5 mM reduced glutathione (GSH) (Merck-Millipore, Burlington, MA) for 24 hours at 37°C.
6
Quantification of Thiol Compounds
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Methanol, acetonitrile (ACN) (HPLC gradient grade, Fisher Scientific, Hampton, NH, USA), trifluoroacetic acid (TFA), ammonium acetate (Sigma-Aldrich, St. Louis, MO, USA) and mili-Q water (Millipore, Milford, MA, USA) were used for the preparation of mobile phases.
L-Cysteine (Cys, 99%), DL-homocysteine (HCys, 95%) and N-acetylcysteine (NAC, ≥99%) were purchased from Sigma-Aldrich (St. Louis, MO, USA), whereas methionine (Met, 99%) and reduced glutathione (GSH, 98%) were supplied by Merck (Darmstadt, Germany). L (+)-ascorbic acid (AA, ≥99%) was obtained from Riedel-de Haën (Seelze, Germany). Chemical structures of these compounds are shown in Scheme 1. 10 mmol L -1 standard stock solutions were prepared in Milli-Q water (Millipore, Milford, MA, USA) and stored at 4 ᵒC. Diluted standard solutions were prepared daily in H2O:ACN (1:1). Sulphuric acid (Merck, Darmstadt, Germany) was used for electrode activation.
L-Cysteine (Cys, 99%), DL-homocysteine (HCys, 95%) and N-acetylcysteine (NAC, ≥99%) were purchased from Sigma-Aldrich (St. Louis, MO, USA), whereas methionine (Met, 99%) and reduced glutathione (GSH, 98%) were supplied by Merck (Darmstadt, Germany). L (+)-ascorbic acid (AA, ≥99%) was obtained from Riedel-de Haën (Seelze, Germany). Chemical structures of these compounds are shown in Scheme 1. 10 mmol L -1 standard stock solutions were prepared in Milli-Q water (Millipore, Milford, MA, USA) and stored at 4 ᵒC. Diluted standard solutions were prepared daily in H2O:ACN (1:1). Sulphuric acid (Merck, Darmstadt, Germany) was used for electrode activation.
7
Comprehensive Biochemical Assays for Oxidative Stress
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Dexamethasone (Dex), aluminum chloride, folin–ciocalteau reagent, 2,2-diphenyl-1-picrylhydrazyl (DPPH), sodium nitroprusside, Griess reagent (sulphanilamide and N-(1-Naphthyl) ethylenediamine dihydrochloride), tris-HCL, pyrogallol, thiobarbituric acid, trichloroacetic acid, 5,5-dithio-bis-2-nitrobenzoic acid (DTNB), cumene H2O2, reduced glutathione (GSH), and phenazine methosulfate fluoride (PMSF) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The total protein assay kit was purchased from BioMED (Cairo, Egypt). Total RNA extraction kit, cDNA synthesis kit, and SYBR green with low ROX were purchased from Enzynomics (Daejeon, Republic of Korea). Primers and antibodies were obtained from Willowfort (Nottingham, UK) and Cell Signaling Technology (Danvers, Massachusetts, USA), respectively. Other chemicals were of analytical grades.
8
Biochemical Assays for Liver and Metabolic Markers
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Alanine aminotransferase (ALT), alkaline phosphatase (ALP), aspartate aminotransferase (AST), total protein, albumin, urea, creatinine, total bilirubin, triglycerides, total cholesterol assay kits, and GOD-POD glucose analysis kit were purchased from Aggappe Diagnostics, Ernakulam, India. Reduced glutathione (GSH) and 5,5-dithio(bis)nitrobenzoic acid (DTNB) were purchased from Sigma-Aldrich, Bangalore, India. All the chemicals and reagents used in the study were of analytical grade.
9
Purification and Characterization of Pyocyanin
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Crystalline pyocyanin was purchased from Cayman or prepared by photolysis of 2 mM solution of phenazine methosulfate in 20 mM TRIS buffer, pH 7.4, as reported previously (Cheluvappa, 2014 (link)); the crude synthetic pyocyanin was recrystallized from the TRIS buffer and judged essentially pure by its UV–Vis and ESI mass-spectra. Reduced glutathione (GSH) was obtained from Sigma. The GSH and GSSG content in glutathione stock solutions was determined fluorimetrically with o-phthaladehyde (Senft et al., 2000 (link)). In-house nanopure water was generated by double distillation and used in all experiments. Stock HEPES buffer (200 mM) was treated with 5% (v/v) Chelex resin (BioRad) by stirring overnight. All other commercial reagents, solvents and media were used without any further purification.
10
Cellular Stress Response Protocol
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3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), paclitaxel (PTX), bafilomycin A1 (BAF-A1), N-acetyl-L-cysteine (NAC), cycloheximide (CHX), catalase-polyethylene glycol (PEG), 4,6-diamidino-2-phenylindole (DAPI), and reduced glutathione (GSH) were purchased from Sigma-Aldrich. 3-Methyladenine (3-MA) and Mn(III) tetrakis (4-benzoic acid) porphyrin chloride (MnTBAP) were purchased from Merck Millipore. ERK/MEK1 inhibitor (selumetinib) was purchased from Selleck Chemicals. DMEM, FBS, and the antibiotics mixture (penicillin-streptomycin) were purchased from Invitrogen. Primary antibodies against β-actin (Chemicon, Millipore), p62 (BD Biosciences), LC3A/B and Ki-67 (Abcam), LC3B (Arigo Biolaboratories.), Raf1 (GeneTex), PDI, caspase-7, Bim, phospho-eIF2α, phospho-ERK1/2, phospho-SAPK/JNK, phospho-p38 MAPK (Cell Signaling Technology) were used. All other antibodies were obtained from Santa Cruz Biotechnology.
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