N acetylcysteine

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom, Sao Tome and Principe, Switzerland, Macao, Italy, China, France, Japan, Spain, Canada, Sweden

N-acetylcysteine is a chemical compound that is commonly used in laboratory settings. It is a precursor to the antioxidant glutathione and has various applications in research and analysis.

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Lab products found in correlation

924 protocols using n acetylcysteine

Detailed Enteroid Culture Protocol

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All enteroid media were prepared as reported previously (32 (link)). Advanced Dulbecco’s modified Eagle’s medium (DMEM)–F-12 medium supplemented with 1× GlutaMAX (Gibco), 10 mM HEPES (Quality Biologicals), and 100 Units/ml penicillin-streptomycin (Gibco) was used as the basal medium.
Complete medium with growth factor (CMGF⁺) is comprised of 22.3% (vol/vol) basal medium, 50% (vol/vol) Wnt3a-conditioned medium, 15% (vol/vol) R-spondin-1-conditioned medium, 10% (vol/vol) noggin-conditioned medium, 1× B27 supplement (Gibco), 1 mM N-acetylcysteine (Sigma), 1× Primocin (InvivoGen), 50 ng/ml human epidermal growth factor (R&D Systems), 10 nM [Leu-15]-gastrin (AnaSpec), 500 nM A83-01 (Tocris), and 10 μM SB202190 (Tocris).
Differentiation medium is comprised of 86.2% (vol/vol) basal medium, 10% (vol/vol) noggin-conditioned medium, 1 mM N-acetylcysteine (Sigma), 50 ng/ml human epidermal growth factor (R&D Systems), 10 nM [Leu-15]-gastrin (AnaSpec), and 500 nM A83-01 (Tocris).

Ranganathan S., Doucet M., Grassel C.L., Delaine-Elias B., Zachos N.C, & Barry E.M. (2019). Evaluating Shigella flexneri Pathogenesis in the Human Enteroid Model. Infection and Immunity, 87(4), e00740-18.

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Cytotoxicity Assay for Cell Viability

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Compounds were added 24 hours after cell plating and viability was assessed at the specified time point using the CellTiter-Glo luminescent cell viability assay (Promega) according to the manufacturer’s instructions. Data were normalized to vehicle-treated controls and IC₅₀ curves were generated using Graphpad Prism. Viability assays were generally performed at least twice. In assays that included N-acetylcysteine (Sigma-Aldrich) treatment, the medium was replaced every 48 hours except in RNAi experiments where additional N-acetylcysteine was added every 48 hours. CB-839 was obtained from Focus Biomolecules (14 (link)). Erastin was purchased from Cayman Chemical (15 (link)), and BPTES was acquired from Sigma (16 (link)).

Beatty A., Fink L.S., Singh T., Strigun A., Peter E., Ferrer C.M., Nicolas E., Cai K.Q., Moran T.P., Reginato M.J., Rennefahrt U, & Peterson J.R. (2017). Metabolite profiling reveals the glutathione biosynthetic pathway as a therapeutic target in triple negative breast cancer. Molecular cancer therapeutics, 17(1), 264-275.

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Optimized Cell Culture Conditions

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All media were prepared as reported previously^{57 (link)}. Advanced Dulbecco’s modified Eagle’s medium (DMEM)–F-12 medium supplemented with 1× GlutaMAX (Gibco), 10 mM HEPES (Sigma-Aldrich), and 100 Units/ml penicillin-streptomycin (Sigma-Aldrich) was used as the basal medium. Expansion medium (EM) is basal medium supplemented with 50% (vol/vol) Wnt3A-conditioned medium, 20% (vol/vol) R-spondin-1-conditioned medium, 10% (vol/vol) Noggin-conditioned medium, 1× B27 supplement (Gibco), 1 mM N-acetylcysteine (Sigma), 1× Primocin (InvivoGen), 50 ng/ml human epidermal growth factor (R&D Systems), 10 nM [Leu-15]-gastrin (AnaSpec), 500 nM A83-01 (Tocris), and 10 μM SB202190 (Tocris). Differentiation medium is comprised of basal medium (no penicillin-streptomycin added), 10% (vol/vol) Noggin-conditioned medium, 1 mM N-acetylcysteine (Sigma), 50 ng/ml human epidermal growth factor (R&D Systems), 10 nM [Leu-15]-gastrin (AnaSpec), 500 nM A83-01 (Tocris), and 10% fetal bovine serum (Sigma Aldrich). Wnt3A (American Type Culture Collection, Manassas, VA), R-spondin1 (kindly provided by Dr. Calvin Kuo, Stanford University, Stanford, CA), and Noggin^{58 (link)} (kindly provided by Dr. Marcel Bijvelds, Erasmus University, Rotterdam, the Netherlands) cell lines were maintained to produce conditioned media.

Liu L., Saitz-Rojas W., Smith R., Gonyar L., In J.G., Kovbasnjuk O., Zachos N.C., Donowitz M., Nataro J.P, & Ruiz-Perez F. (2020). Mucus layer modeling of human colonoids during infection with enteroaggragative E. coli. Scientific Reports, 10, 10533.

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Dietary Manipulations in Murine NASH

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Male wild-type (WT) mice C57Bl/6 were obtained from Jackson Laboratory and fed either chow diet (Picolab^® Rodent diet 20, #5053), methionine-choline deficient (MCD) diet (MP Biomedicals, #960439) for 8 weeks or Western diet (45% high-fat diet, enriched in saturated fat, supplemented with 0.2% cholesterol and high-corn syrup equivalent in the drinking water, TD.120330 22% HVO + 0.2% cholesterol diet, Teklad Research) for 16 weeks. Diet specifications are summarized in Supplemental Tables 1 and 2. Two separate experiments were done, the first compared chow-fed diet (n=6), MCD diet-fed (n=8) and Western diet-fed (n=8) mice; the second experiment studied 30 mice divided into 3 groups: chow diet, MCD diet and MCD diet treated with Pantothenate and N-acetylcysteine. Pantothenate (Sigma C8731) was administered IP in a solution in PBS, at a dose of 250 mg/kg three times per week. The control group was injected with same volume of PBS. N-acetylcysteine (Sigma, A9165) was administered in the drinking water, resulting in an estimated consumption of 250 mg N-acetylcysteine /kg body weight per day.
Animal care and procedures were approved by the Duke University Institutional Animal Care and fulfilled National Institutes for Health and Duke University IACUC requirements for humane animal care.

Machado M.V., Kruger L., Jewell M.L., Michelotti G.A., de Almeida Pereira T., Xie G., Moylan C.A, & Diehl A.M. (2015). Vitamin B5 and N-acetylcysteine in nonalcoholic steatohepatitis: a pre-clinical study in a dietary mouse model. Digestive diseases and sciences, 61(1), 137-148.

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Isolation and Activation of NK Cells

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Splenocytes were isolated and cultured in IL-15 (15 ng/ml; PeproTech) at 37°C for 6 days. On day 4, the cells were supplemented with IL-15 (15 ng/ml) and cultured for an additional 2 d. On day 6, cultured NK cells were magnetically purified (NK cell Isolation Kit II, Miltenyi) and stimulated for 18 hr with IL-2 (20 ng/ml; National Cancer Institute Preclinical Repository) and/or IL-12 (10 ng/ml; Miltenyi Biotec) cytokines. Low-dose IL-15 (6.66 ng/ml) was added as a survival factor to unstimulated cultures. Experiments were carried out in the presence or absence of TEPP-46 (Cayman Chemical or EMD Millipore), rapamycin (20 nM; Fisher), N-acetyl-cysteine (7.5 mM; EMD Millipore) or dehydroepiandrosterone (DHEA) (75 μM, Sigma Aldrich). Splenocytes were cultured in RPMI medium containing 10% FBS, 2 mM glutamine (Thermo Fisher), 50 μM 2-ME (Sigma-Aldrich), and 1% Penicillin/Streptomycin (Thermo Fisher).

Walls J.F., Subleski J.J., Palmieri E.M., Gonzalez-Cotto M., Gardiner C.M., McVicar D.W, & Finlay D.K. (2020). Metabolic but not transcriptional regulation by PKM2 is important for natural killer cell responses. eLife, 9, e59166.

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Carbon Steel Corrosion Inhibition by N-acetylcysteine

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The carbon steel plates, with an active area of 1.0 cm², were submitted to corrosion at room temperature (24 ± 1 °C) and a static regime, in 1.0 mol L⁻¹ HCl blank solution and 1.0 mol L⁻¹ HCl blank solution containing various concentrations of N-acetylcysteine (NAC): 1.5 mmol L⁻¹; 3.0 mmol L⁻¹; 4.5 mmol L⁻¹; 6 mmol L⁻¹. N-acetylcysteine, a Merck product (assay ≥ 99%), is a light-yellow powder and water-soluble. Low-carbon steel (OL 37 Romanian type) was used, containing: C < 0.22; Mn < 0.36%; Si < 0.35%; P < 0.06%; S < 0.06%%; the remainder being Fe up to 100%. Analytical purity hydrochloric acid from Merck and bi-distilled water were used to prepare the uninhibited and inhibited NAC solutions. Before corrosion, the samples were mechanically polished with emery paper, degreased with acetone and dried in warm air.

Samide A., Dobriţescu A., Tigae C., Spînu C.I, & Oprea B. (2023). Experimental and Computational Study on Inhibitory Effect and Adsorption Properties of N-Acetylcysteine Amino Acid in Acid Environment. Molecules, 28(19), 6799.

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Colorectal Cancer Organoid Generation

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Patient-derived colorectal cancer organoids were generated and cultured as described earlier [21 (link), 22 (link)]. In brief, the tissue was cut into small pieces and dissociated at 37 °C. Dissociated cells were passed through a 30 and 100 μm cell strainer and collected in advanced DMEM/F12 medium (Thermo Fisher Scientific GmbH, Waltham, Massachusetts, US) supplemented with Glutamax (Thermo Fisher Scientific GmbH, Waltham, Massachusetts, US), penicillin/streptomycin, HEPES, N-acetylcysteine (Merck Chemicals GmbH, Darmstadt, Germany), N-2 supplement (Thermo Fisher Scientific GmbH, Waltham, Massachusetts, US), B-27 supplement (Thermo Fisher Scientific GmbH, Waltham, Massachusetts, US), EGF (PeproTech GmbH, Hamburg, Germany), Y27632 (Absource Diagnostics GmbH, Munich, Germany), and amphotericin (Sigma-Aldrich Chemical, St. Louis, Missouri, US) and embedded in matrigel (Corning B.V., Amsterdam, Netherlands). For subcultivation, organoids were removed from matrigel and dissociated into small organoids using TrypLE (Thermo Fisher Scientific GmbH, Waltham, Massachusetts, US) and then transferred into fresh matrigel. For coculture experiments, organoids were dissociated into small organoids and 2,500–5,000 small organoids were seeded onto the peritoneum.

Mönch D., Koch J., Maaß A., Janssen N., Mürdter T., Renner P., Fallier-Becker P., Solaß W., Schwab M., Dahlke M.H., Schlitt H.J, & Leibold T. (2021). A human ex vivo coculture model to investigate peritoneal metastasis and innovative treatment options. Pleura and Peritoneum, 6(3), 121-129.

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Directed Differentiation of iPICs for Implantation

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Stage 7 medium was based on a previous report [7 (link)] with our original modifications. Cells were cultured in the MCDB 131 medium with 1% P/S, 2% fat-free bovine serum albumin (FUJIFILM Wako), 20 mM glucose, 1.5 g/L NaHCO₃, 1% GlutaMAX, 0.5% ITS-X (Thermo Fisher Scientific), 10 μM ALK5iII or the alternative candidates, 1 μM T₃, 10 μM ZnSO₄ (Merck Millipore), 1.4 IU/mL heparin sodium salt (Nacalai Tesque, Kyoto, Japan), 1 mM N-acetyl cysteine (Merck Millipore), 10 μM Trolox (FUJIFILM Wako), 2 μM R428 (Selleck Chemicals, Houston, TX, USA), 1 μM PD-166866, 3 μM TR06141363 (Multi-kinase inhibitor, Takeda Pharmaceutical Company), and 10 μM Y-27632, for 4 days. To generate iPICs for implantation, cells were dissociated and re-sized in an Elplasia 6-well microwell plate (Corning Incorporated, Corning, NY, USA) or a gas-permeable microwell culture bag (Toyo Seikan Group Holdings, Ltd., Yokohama, Japan) [20 (link)] and cultured in the stage 7 medium.

Sakuma K., Tsubooka-Yamazoe N., Hashimoto K., Sakai N., Asano S., Watanabe-Matsumoto S., Watanabe T., Saito B., Matsumoto H., Ueno H., Ito R, & Toyoda T. (2023). CDK8/19 inhibition plays an important role in pancreatic β-cell induction from human iPSCs. Stem Cell Research & Therapy, 14, 1.

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Evaluating Novel Compounds for Cellular Treatments

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The compounds used throughout this study and their sources (in parentheses) were the following: ARINA-1 (kindly donated by Renovion, Inc.); camostat mesylate (Millipore Sigma Cas. No. 59721-29-8); poly-N(acetyl, arginyl) glucosamine (PAAG, kindly donated by Synedgen, Inc.); allopurinol (TCI, Cat. No. A0907); ascorbic acid (LETCO Medical, Cat. No. 684471); reduced glutathione (Sigma, Cat. No. G6013); sodium bicarbonate (LETCO Medical, Cat. No. 685940); N-acetylcysteine (EMD Millipore, Cat. No. 106425); L-; BAPTA/AM (EMD Millipore, Cat. No. 196419); sulforaphane (Medkoo Biosciences Inc. Cat. No. 202713250MG). Hydrosoluble and hydrophobic compounds were dissolved in sterile milli-Q water and dimethyl sulfoxide (DMSO), respectively. They were used at the indicated concentration depending on the experiment.

Campos-Gómez J., Fernandez Petty C., Mazur M., Tang L., Solomon G.M., Joseph R., Li Q., Peabody Lever J.E., Hussain S.S., Harrod K.S., Onuoha E.E., Kim H, & Rowe S.M. (2023). Mucociliary clearance augmenting drugs block SARS-CoV-2 replication in human airway epithelial cells. American Journal of Physiology - Lung Cellular and Molecular Physiology, 324(4), L493-L506.

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Nrf2 Regulation by Oxidative Stress

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DFX was purchased from MedChemExpress (Monmouth Junction, NJ, USA). Ammonium ferric citrate (AFC) and N-acetyl-cysteine (NAC) were obtained from Merck Millipore (Burlington, MA, USA). NRF2 and heme oxygenase-1 (HO-1) antibodies were purchased from Genetex (GTX103322, GTX101147; Irvine, CA, USA). KEAP-1 and Ser344 phosphorylation on NRF2 (abbreviated as Phos-Ser344) antibodies were obtained from Thermofisher (PA5-99434, PA5-67520; Waltham, MA, USA). Lys599 acetylation on NRF2 (abbreviated as Ace-Lys599) antibody was purchased from Abbkine (ABP50139; Atlanta, GA, USA). B-Actin (A2066), erastin (571203-78-6), and ferrostatin-1 (347174-05-4) antibodies were purchased from Sigma–Aldrich. HRP Goat antimouse and antirabbit IgG antibodies (Allbio, Taichung, Taiwan) were used as secondary antibodies for western blot analysis. BIO (GSK-3 Inhibitor), EX-527 (SIRT1 inhibitor), and C646 (p300/CBP inhibitor) were purchased from Selleckchem (S7915, S1541, S7152; Houston, TX, USA).

Hsu W.Y., Wang L.T., Lin P.C., Liao Y.M., Hsu S.H, & Chiou S.S. (2024). Deferasirox Causes Leukaemia Cell Death through Nrf2-Induced Ferroptosis. Antioxidants, 13(4), 424.

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