N acetyl l cysteine nac

Manufactured by Fujifilm

Sourced in Japan

N-acetyl-L-cysteine (NAC) is a chemical compound used in various laboratory applications. It is a precursor to the antioxidant glutathione and has been studied for its potential therapeutic properties. NAC serves as a source of sulfhydryl groups and can be used in various biochemical assays and procedures.

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Lab products found in correlation

Mk801, by Fujifilm (2 mentions) S 4 carboxyphenylglycine cpg, by Bio-Techne (2 mentions) Monoclonal rat anti cd68 antibody clone fa 11, by Bio-Rad (2 mentions) Anti ly 6g antibody clone rea526, by Miltenyi Biotec (2 mentions) Diphenyleneiodonium chloride dpi, by Cayman Chemical (2 mentions) L leucyl l leucine methyl ester hydrochloride, by Cayman Chemical (2 mentions) Hoechst 33342, by Dojindo Laboratories (2 mentions) Gp91ds tat, by AnaSpec (2 mentions) Ly341495, by Bio-Techne (1 mentions) Rs α cyclopropyl 4 phosphonophenyl glycine, by Bio-Techne (1 mentions) Metyrapone, by Merck Group (1 mentions) 6 well culture plate, by Corning (1 mentions) Human serum albumin (hsa), by Fujifilm (1 mentions) Dnase 1, by Takara Bio (1 mentions) Bovine serum albumin bsa, by Fujifilm (1 mentions) Sytox green, by Thermo Fisher Scientific (1 mentions) Enspire, by PerkinElmer (1 mentions) Toxinsensor endotoxin assay kit, by GenScript (1 mentions) 6 formylindolo 3 2 b carbazole ficz, by Merck Group (1 mentions) Bortezomib, by Fujifilm (1 mentions)

12 protocols using n acetyl l cysteine nac

Pharmacological Modulation of Neuronal Signaling

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NMDAR antagonist, MK801,15 (link) memantine (MEM), cysteine prodrug, N‐acetyl‐l‐cysteine (NAC),16 (link) and the GABA_A receptor agonist, muscimol (MUS)15 (link) were obtained from Wako Chemicals (Osaka, Japan). The II‐mGluR antagonist LY341495,17 (link) the III‐mGluR antagonist (RS)‐α‐cyclopropyl‐4‐phosphonophenyl glycine (CPPG),17 (link) and the Sxc inhibitor (S)‐4‐carboxyphenylglycine (CPG)18 (link) were purchased from Tocris Bioscience (Bristol, UK).
All compounds were prepared on the day of experiments. MK801, MEM, CPPG, CPG, NAC, and MUS were dissolved in modified Ringer's solution (MRS) or artificial cerebrospinal fluid (ACSF). LY341495 was initially dissolved in 10 mmol L⁻¹ dimethyl sulfoxide and was then diluted to 1 mmol L⁻¹ in MRS. The final concentrations of LY341495 and dimethyl sulfoxide were 1 μmol L⁻¹ and 0.1% (vol vol⁻¹), respectively.
MRS contained 145 mmol L⁻¹ Na⁺, 2.7 mmol L⁻¹ K⁺, 1.2 mmol L⁻¹ Ca²⁺, 1.0 mmol L⁻¹ Mg²⁺, and 154.4 mmol L⁻¹ Cl⁻. The pH was adjusted to 7.4 using 2 mmol L⁻¹ phosphate buffer and 1.1 mmol L⁻¹ Tris buffer.19 (link), 20 (link) ACSF contained 130 mmol L⁻¹ NaCl, 5.4 mmol L⁻¹ KCl, 1.8 mmol L⁻¹ CaCl₂, 1 mmol L⁻¹ MgCl₂, and 5.5 mmol L⁻¹ glucose and was buffered to pH 7.3 using 20 mmol L⁻¹ HEPES.

Okada M., Fukuyama K., Kawano Y., Shiroyama T, & Ueda Y. (2019). Memantine protects thalamocortical hyper‐glutamatergic transmission induced by NMDA receptor antagonism via activation of system xc−. Pharmacology Research & Perspectives, 7(1), e00457.

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Oxidative Stress Masculinization in Medaka

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To confirm the masculinization of XX medaka by oxidative stress, H₂O₂ treatments were performed with 0.75 and 2 mM H₂O₂ (purity 30%, CAS RN: 7722-84-1; Wako Pure Chemical, Osaka, Japan) using wild-type larvae from 0 to 5 days post-hatching (dph) in 6-well culture plates (Corning, Glendale, AZ) with the water being changed daily. A rescue test to reduce oxidative stress through antioxidant supplementation was conducted with either 1 or 10 μM N-acetyl-L-cysteine (NAC; Wako) dissolved in Dimethyl sulfoxide (DMSO; Sigma-Aldrich, Saint Louis, MO) as previously described (20 (link)), using larvae from 0 to 5 dph in 6-well culture plates with the water being changed daily.
To investigate whether other XX medaka lines undergo masculinization from oxidative stress, pparaa KO, and gsdf KO medaka larvae were treated with 2 mM H₂O₂ under the conditions outlined above. Finally, treatment with 5 μM metyrapone (Sigma-Aldrich) dissolved in ethanol (Wako) as previously described (17 (link)), was used in conjunction with 2 mM H₂O₂ in FLFII medaka larvae to investigate the relationship between cortisol and oxidative stress-induced masculinization. The survival rates and the body sizes in adults are shown in Supplementary Tables S1, S2, respectively.

Mukai K., Hara S., Sakima K., Nozu R., Yazawa T, & Kitano T. (2022). Oxidative Stress Causes Masculinization of Genetically Female Medaka Without Elevating Cortisol. Frontiers in Endocrinology, 13, 878286.

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Extracellular DNA Quantification Protocol

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Extracellular DNA was stained with 5 μm SYTOX Green (Life Technologies, Carlsbad, CA, USA), a fluorescent membrane‐impermeable DNA dye. Fluorescence was quantified using a microplate reader equipped with filters to detect excitation/emission maxima of 504/523 nm (EnSpire; PerkinElmer, Waltham, MA, USA). When indicated, neutrophils were stimulated with 20 or 200 nm PMA, or cultured in the presence of 100 U·mL⁻¹ DNase I (TaKaRa, Osaka, Japan), 40 mg·mL⁻¹ bovine serum albumin (BSA; Wako Pure Chemical Industries, Osaka, Japan), 40 mg·mL⁻¹ human serum albumin (Wako), heat‐treated (complement‐inactivated) serum, protein G/A sepharose‐treated (immunoglobulin‐depleted) serum, 10 mm EDTA, or 5 or 20 mm N‐acetyl‐l‐cysteine (NAC; Wako).

Kamoshida G., Kikuchi‐Ueda T., Nishida S., Tansho‐Nagakawa S., Kikuchi H., Ubagai T, & Ono Y. (2017). Spontaneous formation of neutrophil extracellular traps in serum‐free culture conditions. FEBS Open Bio, 7(6), 877-886.

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Pharmacological Modulation of Glutamate Signaling

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The NMDA-R antagonists, MK801 and amantadine (AMA) [17 (link)], and a cysteine prodrug, N-acetyl-L-cysteine (NAC) [18 (link)] were obtained from Wako Chemicals (Osaka, Japan). The cystine/glutamate antiporter (Sxc) inhibitor (S)-4-carboxyphenylglycine (CPG) [19 (link)] was purchased from Tocris Bioscience (Bristol, UK). Selective group I metabotropic glutamate receptors (I-mGluRs) antagonist, YM298198, was purchased from Sigma-Aldrich (St. Louis, MO, USA). A selective AMPA-R antagonist, perampanel (PER), was obtained from Cosmo Bio (Tokyo, Japan). All compounds were prepared on the day of their use in experiments. MK801, AMA, YM298198 and NAC were dissolved in modified Ringer’s solution (MRS) or artificial cerebrospinal fluid (ACSF). PER was initially dissolved at a concentration of 1 mM in dimethyl sulfoxide. CPG was initially dissolved in 1 N NaOH and was diluted to 1 μM in MRS.

Nakano T., Hasegawa T., Suzuki D., Motomura E, & Okada M. (2019). Amantadine Combines Astroglial System Xc− Activation with Glutamate/NMDA Receptor Inhibition. Biomolecules, 9(5), 191.

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Silica Nanoparticles and Microparticles Characterization

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Amorphous silica nanoparticles (50 nm diameter without surface modification; 50 nm-plain with amine surface modification; 50 nm-NH₂) and microparticles (3 μm diameter; 3 μm-plain) used in this study were purchased from Micromod Partikeltechnologie GmbH (Rostock, Germany). In some experiments, we used nanoparticles labeled with FITC for their localization. All the products contained no detectable endotoxin as assessed by a ToxinSensor Endotoxin Assay kit (GenScript, Piscataway, USA). Prior to in vitro and in vivo studies, the particles were vortexed for 60 s.
Monoclonal rat anti-CD68 antibody (Clone FA-11) was purchased from Bio-rad (Hercules, CA, USA). Anti-F4/80 antibody (CI: A3–1) was from (Caltag Laboratories, Burlingame, CA, USA). Anti-Ly-6G-antibody (Clone REA526) was from Miltenyi Biotec (Bergisch Gladbach, Germany). Hoechst33342 was from Dojindo (Kumamoto, Japan). L-leucyl-L-leucine methyl ester (hydrochloride) was from Cayman Chemical (Ann Arbor, MI, USA). N-acetyl-L-cysteine (NAC) was from Wako Pure Chemicals (Osaka, Japan). Diphenyleneiodonium chloride (DPI) was from Cayman Chemical. gp91ds-tat was from Anaspec (Fremont, CA, USA).

Inoue M., Sakamoto K., Suzuki A., Nakai S., Ando A., Shiraki Y., Nakahara Y., Omura M., Enomoto A., Nakase I., Sawada M, & Hashimoto N. (2021). Size and surface modification of silica nanoparticles affect the severity of lung toxicity by modulating endosomal ROS generation in macrophages. Particle and Fibre Toxicology, 18, 21.

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PM2.5-Induced Oxidative Stress in A549 Cells

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A549 cells were cultured in Dulbecco’s modified eagle’s medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C with 5% CO₂. PM_2.5 suspension was applied to A549 cells at a final concentration of 2 µg/mL for four days. BaP (Tokyo Chemical Industry, Tokyo, Japan) and 6-Formylindolo[3,2-b] carbazole (FICZ, Sigma-Aldrich, St. Louis, MO) were dissolved in dimethyl sulfoxide (DMSO) and stored at 4 mM concentration. A549 cells were seeded into 6-well plates at 5×10⁴ cells concentration and treated with BaP or FICZ at the indicated concentrations and times. Control cells were treated with DMSO alone. For H₂O₂ stimulation, A549 cells were seeded into 6- or 12-well plates at 2×10⁵ or 5×10⁵ cells, respectively, and treated with 10 µM H₂O₂ (Nacalai Tesque, Tokyo, Japan) for 6 h. N-acetyl-l-cysteine (NAC, FUJIFILM Wako Chemicals, Tokyo, Japan) was reapplied to A549 cells at a final concentration of 1 mM for 1 h with 2 µM BaP.

Kohno R., Nagata Y., Ishihara T., Amma C., Inomata Y., Seto T, & Suzuki R. (2023). Benzo[a]pyrene induces NLRP1 expression and promotes prolonged inflammasome signaling. Frontiers in Immunology, 14, 1154857.

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Bortezomib and NAC Synergistic Treatment

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Bortezomib (021–18,901) and N‐acetyl‐l‐cysteine (NAC) (017–05131) were purchased from FUJIFILM Wako Pure Chemicals. Phorbol 12‐Myristate 13‐Acetate (P8139) was purchased from Sigma‐Aldrich.

Abe K., Ikeda S., Nara M., Kitadate A., Tagawa H, & Takahashi N. (2023). Hypoxia‐induced oxidative stress promotes therapy resistance via upregulation of heme oxygenase‐1 in multiple myeloma. Cancer Medicine, 12(8), 9709-9722.

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Quantification of Cellular GSH and ROS

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Cells were seeded in dark-colored, flat-bottomed 96-well plates. In GSH assays, cells at a density of 5 x 10³ cells/well were treated with GSH-Glo reagent (Promega, Madison, WI, USA) as stipulated by the manufacturer and luminescence was analyzed in a microplate reader. In ROS assays, cells at a density of 2 x 10⁴ cells/well were stained with 10 μM of dichlorodihydrofluorescein diacetate (DCF-DA) (Sigma-Aldrich, St. Louis, MO, USA) for 45 minutes, treated with a ROS inducer (5 μM of CDDP) and/or ROS scavenger [5 or 10 mM of N-Acetyl-L-cysteine (NAC) (Wako, Osaka, Japan)] for 4 hours, and then analyzed in a microplate reader. We utilized a previously described protocol for GSH and ROS analyses [18] (link). Data was representative of three independent experiments with 4 replicates.

Mvunta D.H., Miyamoto T., Asaka R., Yamada Y., Ando H., Higuchi S., Ida K., Kashima H, & Shiozawa T. (2017). SIRT1 Regulates the Chemoresistance and Invasiveness of Ovarian Carcinoma Cells. Translational Oncology, 10(4), 621-631.

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Intracellular ROS Measurement Assay

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To assess intracellular ROS production, cells were seeded in 96-well plates at a density of 1 × 10⁵ cells/mL and into 60 mm dish (1 × 10⁵ cells/mL). After exposure to butein and CORT, 10 μM Dichloro-dihydro-fluorescein diacetate (DCFH-DA, Sigma Aldrich, USA) was added and cells were incubated for 1 h at 37 °C. Cells were washed twice with HBSS (Fujifilm Wako, Japan) and changes in fluorescence intensity of DCFH-DA were immediately measured using a fluorescence plate reader (Tecan, Switzerland) with excitation at 475 nm and emission at 525 nm. In imaging analysis, nuclei were stained with Hoechst 33342 and observed under a fluorescence microscope (Carl Zeiss, Germany). The ROS generation was evaluated by fluorescence of 2′, 7′-Dichlorodihydrofluorescein (DCF) in combination with N-acetyl-L-cysteine (NAC) (Fujifilm Wako, Japan). NAC is a well-known ROS scavenger, and at final concentration of 50 μM, it was used to identify whether CORT induces or inhibits ROS production.

Ohmoto M., Shibuya Y., Taniguchi S., Nakade T., Nomura M., Ikeda-Matsuo Y, & Daikoku T. (2020). Protective effects of butein on corticosterone-induced cytotoxicity in Neuro2A cells. IBRO Reports, 8, 82-90.

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Oxidative Stress Modulation in MDA-MB-231 Cells

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The MDA-MB-231 cells were collected in a buffer with 10% FBS in phosphate-buffered saline (PBS) and transferred in a sterile light-blocking microtube before being stained with 20 µM of 2’,7’-dichlorofluorescein diacetate (DCFDA) (Sigma #D6883, USA) and incubated for 30 minutes. Later, γ-mangostin and α-mangostin were added to the cell suspensions and incubated in a 37ºC incubator (with 5% CO₂) before analyzing by a flow cytometer (Calibur) at indicated times (4, 8, 18, and 24 hours). 2’,7’-dichlorofluorescein diacetate reacted with the hydroxyl radicals (·OH) and transformed into fluorescent 2,7-dichlorofluorescein (DCF). The fluorescence intensity of DCF based on 5,000 cells per gate was calculated using the flow cytometer (29 (link), 30 (link)). In a separate experiment, the cells already stained with DCFDA were treated with 5 mM N-acetyl-L-cysteine (NAC) (Wako #017-05131, Japan) and incubated for an hour before the treatment with γ-mangostin and α-mangostin for the next 24 hours, and the ROS level was subsequently measured. The assay was carried out in triplicate.

Sarmoko S., Novitasari D., Toriyama M., Fareza M.S., Choironi N.A., Itoh H, & Meiyanto E. (2023). Different Modes of Mechanism of Gamma-Mangostin and Alpha-Mangostin to Inhibit Cell Migration of Triple-Negative Breast Cancer Cells Concerning CXCR4 Downregulation and ROS Generation. Iranian Journal of Pharmaceutical Research : IJPR, 22(1), e138856.

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