RAS-selective lethal (RSL3, S8155) and ferrostatin-1 (Fer-1, S7243) were purchased from Selleck Chemicals (Shanghai, China) and dissolved in dimethyl sulfoxide (DMSO, V900090, Sigma-Aldrich). RSL3 and Fer-1 were prepared for intraperitoneal (i.p.) injection as follows: 1% RSL3 or Fer-1 + 30% PEG300 (S6704, Selleck) + 5% Tween 80 (S6702, Selleck) + 64% H2O. N-Acetyl-l -cysteine (NAC, A9165, Sigma-Aldrich) was used. An optimal cutting temperature compound (OCT, 14020108926) was obtained from Leica (Wetzlar, Germany).
N acetyl l cysteine nac a9165
Manufactured by Merck Group
Sourced in United States
N-acetyl-L-cysteine (NAC) (A9165) is a chemical compound that functions as a precursor to the antioxidant glutathione. It is a white crystalline powder that is soluble in water. NAC is commonly used in research and laboratory settings, but its specific intended use is not provided.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Rsl3 s8155, by Selleck Chemicals (1 mentions)
Fer 1 s7243, by Selleck Chemicals (1 mentions)
Fer 1, by Selleck Chemicals (1 mentions)
Peg300, by Selleck Chemicals (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Mouse c2c12 myoblast, by American Type Culture Collection (1 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (1 mentions)
Collagenase type 1, by Merck Group (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Cellrox green, by Thermo Fisher Scientific (1 mentions)
Matrigel, by Corning (1 mentions)
Anti flag m2 affinity gel, by Merck Group (1 mentions)
Rotenone, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Nuclear and cytoplasmic protein extraction kit, by Beyotime (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Sybr green realtime pcr master mix, by Toyobo (1 mentions)
12 protocols using n acetyl l cysteine nac a9165
1
Preparation and application of RAS-selective lethal and ferroptosis inhibitors
2
Myoblast Differentiation and Aldosterone Signaling
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Mouse C2C12 myoblasts were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 15% fetal bovine serum (FBS), 20 mM hydroxyethyl piperazine ethane sulfonic acid (HEPES), 2 mM L-glutamine, 100 U/mL penicillin, and 0.1 mg/mL streptomycin (Life Technologies Corp., Carlsbad, CA, USA) at 37°C in a humidified atmosphere containing 5% CO2. To induce myogenesis, cells were grown to 90% confluency in maintenance media and switched to differentiation media (DMEM with 2% horse serum) for 3 or 4 days of culture. Recombinant aldosterone (A9477), eplerenone (E6657), and N-acetyl-L-cysteine (NAC, A9165) were purchased from Sigma-Aldrich (St. Louis, MO, USA).
3
Culturing Breast Cancer Cell Lines
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MCF-7, MCF-7 with GFP-LC3 (GFP-LC3-MCF-7) (1), and HS578T cells were cultured in RPMI1640 (Gibco, 22400-105) containing 10% fetal bovine serum (FBS; Gibco, 16000-044) and 1% penicillin/streptomycin (Invitrogen, 15140-122) at 37°C in a 5% humidified CO2 incubator. Primary breast cancer tissues were obtained from consenting patients, and the study protocol was approved by the Research Ethics Board at Asan Medical Center. Tumor specimens were minced with scissors and subsequently digested in Minimum Essential Medium (MEM; Gibco, 11095-098) that contained 1 mg/mL type I collagenase (Sigma, C2799) at 37°C for 2 h. Cells were washed with medium containing 10% FBS, followed by a phosphate-buffered saline (PBS, pH 7.4) wash to remove the FBS. Cells were plated in Mammary Epithelial Basal Medium (MEBM; Lonza, cc-3151) and cultured at 37°C in a 5% CO2 incubator. Actinomycin D (ACD; A9415), BIX-01294 (BIX; B9311), caffeic acid phenethyl ester (CAPE; C8221), cyclohexamide (CHX; C1810), 5-Aza-2′-deoxycytidine (5-Aza-Cd; A3656), and N-acetyl-L-cysteine (NAC; A9165) were purchased from Sigma.
4
Microcystin Detection and Cellular Assays
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Microcystins were obtained from Enzo Life Sciences (Farmingdale, NY, USA) and Cayman Chemical Company (Ann Arbor, MI, USA). The Microcystin (Adda specific) ELISA kit was purchased from Enzo Life Sciences. Pranlukast (P0080), thiazolyl blue tetrazolium bromide (M2128), suramin sodium salt (S2671), and N-acetyl-L-cysteine (NAC) (A9165) were from Sigma (Burlington, MA, USA). Rat tail collagen (354236) and Matrigel (354234) were from Corning (Corning, NY, USA). The TUNEL assay kit (C10247) and CellRox Green (C10444) were obtained from Invitrogen (Waltham, MA, USA). A list of antibodies and dyes used in the study can be found in Supporting Table S1 .
5
Mitochondrial Dysfunction Modulation Protocol
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Muscone and (+)-borneol (purity ≥ 98%) were purchased from Chengdu Must Biotechnology Co., Ltd. (Chengdu, China). Rose bengal (330000), 2,3,5-triphenyl tetrazolium chloride (TTC, T8877), Evans blue (E2129), lipopolysaccharide (LPS, L2880), dimethyl malonate (DMM, 136441), dimethyl succinate (W239607) and N-acetyl-L-cysteine (NAC, A9165) were purchased from Sigma (St. Louis, MO, USA), Cyclosporin A (CSA, HY-B0579), calcimycin (HY-N6687) and mito-TEMPO (HY-112879) were obtained from Med Chem Express (Brea, CA, USA). IL-1β (AF-211-11B) was purchased from Peprotech (Cranbury, NJ, USA).
6
Protein Extraction and Analysis Protocol
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N-Acetyl-l -cysteine (NAC, A9165), LPS, rotenone, antimycin A, mg132 (C2211), His beads (P6611) and ANTI-FLAG® M2 Affinity Gel (A2220) were from Sigma-Aldrich. The nuclear and cytoplasmic protein extraction kit was from Beyotime Biotech. Blue Native PAGE Sample Buffer (C506055-0005), native PAGE (C61104-0001) and oligo nucleotides were synthesized by Sangon Biotech. The Phos-tag TM Acrylamide AAL-107 was from the NARD Institute.
7
Molecular Mechanisms of Kaempferide in Cellular Signaling
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Kaempferide was obtained from the TOKYO chemical industry (TCI, Tokyo, Japan). SYBR® Green Realtime PCR Master Mix was obtained from Toyobo (Tokyo, Japan), and TRIzol® Reagent was obtained from Invitrogen (Waltham, MA, USA). Rabbit anti-phospho-MAPKs (ERK: 9102S, p38: 9212S, JNK: 9251S), rabbit anti-MAPKs (ERK: 9101S, p38: 9211S, JNK: 9252S) and rabbit anti-phospho-AKT (4060S) and rabbit anti-AKT (9272S) were obtained from Cell Signalling Technology, Inc. (Danvers, MA, USA). Rabbit anti-COL1A1 (PA5-29569) and rabbit anti-MMP-1a (PA5-27210) were obtained from Invitrogen (Middlesex County, MA, USA). N-acetyl-L-cysteine (NAC) (A9165) was purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).
8
Antibody Validation for BCAA Metabolism
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Antibodies against BCKDHA (sc-271538) and β-actin (sc-47778) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against BCAT1 (12822), BCAT2 (9432), and ACLY (4332) were purchased from Cell Signaling Technology (Danvers, MA, USA). N-acetyl-l -cysteine (NAC; A9165) and palmitic acid (P0500) were obtained from Sigma-Aldrich (St. Louis, MO, USA). AdipoRed assay reagent (#PT-7009) was obtained from Lonza (Basel, Switzerland).
9
ISP-I Dissolution and Characterization
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ISP-I (95.9% purity) was procured from Shanghai Tonglian Pharmaceutical Co., Ltd. (China). For cell experiments, ISP-I was dissolved to 100mM stock concentration in DMSO (Sigma) and diluted to appropriate concentrations with cell culture medium (final DMSO concentration in culture medium was no greater than 0.015%). For animal experiments, ISP-I was dissolved in Polyethylene glycol (PEG; Sigma), vegetable oil and Tween-80 (Sigma) mixed solvents (v:v:v = 9.5:9.5:1) at 24mg/ml, and then diluted to the final concentration with sterilized water.
The CCK-8 solution, DCFH-DA and crystal violet solution were all procured from Beyotime (Shanghai, China). Dihydroethidium (DHE, #D7008) and N-acetyl-l-cysteine (NAC, A9165) were provided by Sigma (MO, USA). The antibodies used in our research were provided in SupplementaryTable S1 .
The CCK-8 solution, DCFH-DA and crystal violet solution were all procured from Beyotime (Shanghai, China). Dihydroethidium (DHE, #D7008) and N-acetyl-l-cysteine (NAC, A9165) were provided by Sigma (MO, USA). The antibodies used in our research were provided in Supplementary
10
Drug Screening Using Inhibitor Libraries
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Libraries containing PPI inhibitor (L9400) and ubiquitination compound (L8600)— for drug screening—were obtained from TargetMol (Boston, MA, USA). Molibresib (10676), I-BET151 (11181) and dBET1 (18044) were obtained from Cayman Chemical Co. (Ann Arbor, MI, USA). N-acetyl-L-cysteine (NAC) (A9165), Bafilomycin A1 (B1793), and KU55933 (SML1109) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Hoechst 33,342 (H3570) was obtained from Thermo Fisher Scientific (Waltham, MA, U.SA.) and DRAQ5 (ab108410) was obtained from Abcam (Cambridge, U.K.). Short interfering RNA (siRNA) targeting BRD4 (#1, 5′-AGAUUGAAAUCGACUUUGAUU-3′ and #2, 5′-UGAGCACAAUCAAGUCUAAUU-3′), NBR1 (5′-GAACGUAUACUUCCCAUUGUU-3′), p62/SQSTM1 (5′-GCAUUGAAGUUGAUAUCGAUUU-3′), and scrambled siRNA (5′-CCUACGCCACCAAUUUCGU-3′) were synthesized by Genolution (Seoul, Korea).
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