L glutathione reduced gsh

Manufactured by Merck Group

Sourced in United States, Germany

L-glutathione reduced (GSH) is a tripeptide composed of glutamic acid, cysteine, and glycine. It is a naturally occurring antioxidant that plays a crucial role in maintaining the redox balance within cells. GSH helps neutralize free radicals and reactive oxygen species, protecting cells from oxidative stress.

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50 protocols using l glutathione reduced gsh

Synthesis of Silver Nanoparticles

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Milli-Q water (18.2 MΩ.cm/25 °C) was used throughout this study. All glassware were washed with 0.05 M nitric acid, rinsed with Milli-Q water, and dried in an oven overnight. AgNO₃, (99.9999%), L-glutathione reduced (GSH) and sodium borohydride (NaBH₄) were purchased from Sigma-Aldrich. L-cysteine (≥ 99%) was purchased from Beijing Solarbio Science & Technology Co., Ltd. The other reagents (analytical reagent) were purchased from China National Pharmaceutical Group Corporation.

Zhang L., Goswami N., Xie J., Zhang B, & He Y. (2017). Unraveling the molecular mechanism of photosynthetic toxicity of highly fluorescent silver nanoclusters to Scenedesmus obliquus. Scientific Reports, 7, 16432.

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Enzymatic Assay for Antioxidant Activity

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β-nicotinamide adenine dinucleotide 2'-phosphate reduced (β-NADPH), ethylenediaminetetraacetic acid (EDTA), glutathione reductase (GSH-R), 4-nitrophenyl-α-D-glucopyranoside, 4-nitrophenyl-N-acetyl-β-D-glucosaminide, and L-glutathione reduced (GSH) were purchased from Sigma-Aldrich Co. LLC. (USA). Deionized water (DW) was prepared with a Milli-Q Water Purifier (Millipore SAS, France). Liquid nitrogen was supplied by a nearby gas plant.

Kang S.M., Kang G., Seong P.N., Park B., Kim D, & Cho S. (2014). Evaluation of the Activities of Antioxidant Enzyme and Lysosomal Enzymes of the Longissimus dorsi Muscle from Hanwoo (Korean Cattle) in Various Freezing Conditions. Korean Journal for Food Science of Animal Resources, 34(6), 742-748.

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Glutathione S-Transferase Activity Assay

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The sample was prepared the same way as for the ChE assay. For the GST assay, we followed the Mannervik method [32 (link)]. A total of 50 µL of the supernatant (100 mM K-P buffer, pH 8 with 0.1% Triton-X 100 for blank) was transferred into a 96 well microtiter plate with 50 µL of 4 mM 1-Chloro-2,4-dinitrobenzene (CDNB) (Sigma Aldrich, St. Louis, MO, USA), prepared in absolute ethanol and 50 µL of 4 mM L-glutathione reduced (GSH) (Sigma-Aldrich, St. Louis, MO, USA), prepared in K-P buffer, pH 8. Absorbance values were measured at 340 nm using a spectrophotometer Cytation 3 (BioTek, Bad Friedrichshall, Germany) for 20 cycles (at 1 min intervals, for 20 min). The activity of GST was expressed as nM/min/mg of proteins, therefore also the concentration of proteins was measured in each sample using the same protocol as described for the ChE assay. All measurements were performed at RT in triplicates. The experiment was performed in three repetitions.

Jan Z., Hočevar M., Kononenko V., Michelini S., Repar N., Caf M., Kocjančič B., Dolinar D., Kralj S., Makovec D., Iglič A., Drobne D., Jenko M, & Kralj-Iglič V. (2023). Inflammatory, Oxidative Stress and Small Cellular Particle Response in HUVEC Induced by Debris from Endoprosthesis Processing. Materials, 16(9), 3287.

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Biliverdin Hydrochloride and Glutathione Rat Study

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Biliverdin hydrochloride (Frontier Scientific, Logan, UT, USA) was dissolved in 0.2 mol/L NaOH, and the solution was adjusted to a final pH of 7.4 with hydrogen chloride. L-glutathione reduced (GSH) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Male Sprague-Dawley rats, weighing 200–230 g, were purchased from Jiangnan University Animal Center and housed in a room with a 12/12-h light/dark cycle. Rats had ad libitum access to food and water. Experiments were carried out according to protocols approved by the Animal Care and Use Committee of Jiangnan University.

Tian W.F., Weng P., Sheng Q., Chen J.L., Zhang P., Zhang J.R., Du B., Wu M.C., Pang Q.F, & Chu J.J. (2017). Biliverdin Protects the Isolated Rat Lungs from Ischemia-reperfusion Injury via Antioxidative, Anti-inflammatory and Anti-apoptotic Effects. Chinese Medical Journal, 130(7), 859-865.

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Synthesis and Characterization of Antimicrobial Nanocomposites

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L-Glutathione reduced (GSH), hydrochloric acid (HCl), sodium nitrite, and polyethylene glycol (MW = 3,350) were purchased from Sigma-Aldrich (St. Louis, MO). Acetone; DOWSIL 3140, MIL-A-46146 RTV Silicone Coating; Dow Corning Silastic Laboratory Tubing (ID 0.058" × OD 0.077", 2415542); and Helix Medical Inc. Silicone Tubing (0.125" × 0.250", 6001121) were purchased from Fisher Scientific Inc. (Pittsburgh, PA). Male Luer Lock Injection Site caps (80149) were purchased from Qosina (Ronkonkoma, NY). Luria-Bertani (LB) agar broth and 10 mM phosphate buffered saline (PBS) (pH 7.2) were purchased from ThermoFisher Scientific (Grand Island, NY). Zinc oxide nanoparticles (APS 30 nm in diameter) were purchased from EPRUI Biotech Co. Ltd. (ShangHai, China). All aqueous solutions were prepared with 18.2 M Ω deionized water using a Milli-Q filter (Milli-q purified water) from EMD Millipore (Billerica, MA). Staphylococcus aureus ATCC 25923 and Pseudomonas aeruginosa ATCC 27853 were obtained from the American Type Culture Collection (Manassas, VA).

Doverspike J.C., Mack S.J., Lou A., Stringer B., Reno S., Cornell M.S., Rojas-Pena A., Wu J., Xi C., Yevzlin A, & Meyerhoff M.E. (2020). Nitric Oxide-Releasing Insert for Disinfecting the Hub Region of Tunnel Dialysis Catheters. ACS applied materials & interfaces, 12(40), 44475-44484.

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Hydrogel Composition and Characterization

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L-Glutathione reduced (GSH) (CAS number 70-18-8) was supplied by Sigma Aldrich (St. Louis, MO, USA). Furcellaran (FUR) (type 7000) was procured from Est-Agar AS (Karla village, Estonia). The chemical content composition of FUR (Mw= 2.951 × 105) was: carbohydrates 79.61%; protein 1.18% and fat 0.24%. Chitosan (CHIT) (molecular weight ~890,000, CAS number 9012-76-4 ≥ 90% deacetylation, 100–300 cP viscosity; particle size ≤ 100 mesh) was supplied by POL-AURA (Zabrze, Poland). Sodium hydroxide (>99%) was purchased from Sigma Aldrich (St. Louis, MO, USA).

Drozdowska M., Piasna-Słupecka E., Such A., Dziadek K., Krzyściak P., Kruk T., Duraczyńska D., Morawska-Tota M, & Jamróz E. (2024). Design and In Vitro Activity of Furcellaran/Chitosan Multilayer Microcapsules for the Delivery of Glutathione and Empty Model Multilayer Microcapsules Based on Polysaccharides. Materials, 17(9), 2047.

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Synthesis and Characterization of Citrate-Stabilized AuNPs

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All of the DNA oligonucleotides were from Integrated DNA Technologies (IDT, Coralville, IA, USA; see Table 1 for sequences). Sodium citrate, potassium cyanide (KCN), and 4-(2-hydroxyethyl) piperazine-1-ethanesulfonic acid (HEPES) were from Mandel Scientific (Guelph, ON, Canada). BSA, FITC-labeled BSA (FITC-BSA), catalase (CAT), hemoglobin (Hb), glucose oxidase (GOx), horseradish peroxidase (HRP), hydroxy-1-hexanethiol (MCH), and L-glutathione reduced (GSH) were from Sigma-Aldrich. The 13 and 24 nm AuNPs were prepared using the citrate reducing method (Liu and Lu, 2006 (link)), and these were called citrate-AuNPs. Milli-Q water was used to prepare all of buffers and solutions.

Wu R., Peng H., Zhu J.J., Jiang L.P, & Liu J. (2020). Attaching DNA to Gold Nanoparticles With a Protein Corona. Frontiers in Chemistry, 8, 121.

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GSK-3β Enzyme Kinetics Assay

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The chemical structure of all of the compounds used in this study is
given in Fig. 1. COB-187 was prepared as
described previously¹⁵.
Tideglusib, alsterpaullone, AR-A014418, hypothemycin, ethanethiol,
tributylphosphine, and THF were purchased from Sigma-Aldrich (St. Louis, MO).
Insect-expressed human recombinant full length histidine-tagged GSK-3β
(catalog number PR5074A) was from Life Technologies/Thermo Fisher (Waltham, MA).
The mutant version of the abovementioned GSK-3β, where the cysteine at
position 199 was replaced with an alanine (C199A), was generated by ThermoFisher
via a contract agreement. The phospho substrate (catalog number AQT0157) and ATP
(catalog number AQT 100XATP) were obtained from AssayQuant Tech (Marlboro, MA).
L-Glutathione reduced (GSH) (catalog number: G4251) and
Tris(2-carboxyethyl)phosphine hydrochlo (TCEP) (catalog number: C4706) were
purchased from Sigma-Aldrich. Dithiotheritol (DTT) (catalog number: AQT1000X
DTT) was purchased from AssayQuant Tech. The kinase reaction buffer (AssayQuant
Tech; catalog number: AQT10XRB) contained 500 mM HEPES, pH 7.5, 0.1% Brij-35,
and 100 mM MgCl₂. The enzyme dilution buffer (AssayQuant Tech;
catalog number: AQT1XEDB) contained 20 mM HEPES, pH 7.5, 0.01% Brij-35, 5%
glycerol, and 1 mg/mL bovine serum albumin.

Ghazanfari D., Noori M.S., Bergmeier S.C., Hines J.V., McCall K.D, & Goetz D.J. (2021). A novel GSK-3 inhibitor binds to GSK-3β via a reversible, time and Cys-199-dependent mechanism. Bioorganic & medicinal chemistry, 40, 116179.

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Mitochondrial Dysfunction and Cell Death

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Chemicals were as follows: Menadione (Sigma Aldrich), 1,4-Benzoquinone (BZQ; USP), 2,3-Dimethoxy-1,4-naphthoquinone (DMNQ, Sigma Aldrich), Mitoxantrone dihydrochloride (MTX, Sigma Aldrich), Carbonyl cyanide 3-chlorophenylhydrazone (CCCP; Tocris), Monobromobimane (MBB; Thermo Fischer Scientific), L-glutathione reduced (GSH; Sigma Aldrich), N-Acetyl-L-cysteine (NAC; Sigma Aldrich), 4′,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI; Thermo Fischer Scientific), Rotenone (Sigma Aldrich), Z-VAD.fmk, (BACHEM); DilC₁ (5) (Thermo Fischer Scientific). ECL chemiluminescence detection kit (GE Healthcare).
Antibodies against the following proteins were used: actin (mouse mAb; CHEMICON); AIF (mouse mAB; Santa Cruz; rabbit pAB; Cell Signaling); Hsp60 (mouse mAb; Stressgen); caspase 3 (rabbit pAB; Cell Signaling); PARP (rabbit pAB; Cell Signaling); VDAC (rabbit pAB; Cell Signaling); Cytochrome C (mAB BD Bioscience).

Wiraswati H.L., Hangen E., Sanz A.B., Lam N.V., Reinhardt C., Sauvat A., Mogha A., Ortiz A., Kroemer G, & Modjtahedi N. (2016). Apoptosis inducing factor (AIF) mediates lethal redox stress induced by menadione. Oncotarget, 7(47), 76496-76507.

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Polyelectrolyte Complex Formation

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Poly(methacrylic acid) (PMAA, average M_w 100,000 Da), poly(N-vinylpyrrolidone) (PVPON, average M_w 1,300,000 Da), poly(ethyleneimine) (PEI; average M_w 70,000 Da), l-glutathione reduced (GSH), and cystamine dihydrochloride (CS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). 1-ethyl-3-(3-(dimethylamino)propyl)-carbodiimide hydrochloride (EDC) was purchased from Chem-Impex International (Wood Dale, IL, USA). Monobasic sodium phosphate (NaH₂PO₄), dibasic sodium phosphate (Na₂HPO₄), ethylenediaminetetraacetic acid sodium salt (EDTA), sodium chloride (NaCl), calcium chloride (CaCl₂), sodium carbonate (Na₂CO₃), ethidium bromide, and calf thymus DNA (ctDNA) were purchased from Fisher Scientific (Hampton, NH, USA). DNA oligonucleotides (5’-AGGGTTAGGGTTAGGGTTAGGG-3’) were purchased from Midland Certified Reagents (Midland, TX, USA).

Alford A., Tucker B., Kozlovskaya V., Chen J., Gupta N., Caviedes R., Gearhart J., Graves D, & Kharlampieva E. (2018). Encapsulation and Ultrasound-Triggered Release of G-Quadruplex DNA in Multilayer Hydrogel Microcapsules. Polymers, 10(12), 1342.

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