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Submit ReviewN-Acetylcysteine
Description: Glutathione (GSH) precursor; maintains stem cell function in culture.
Chemical Name: (2R)-2-Acetamido-3-sulfanylpropanoic acid
Purity: ≥98% (HPLC)
Biological Activity for N-Acetylcysteine
N-Acetylcysteine is a cell-permeable antioxidant and a precursor of reduced glutathione (GSH) (Cat. No. 5219), with anti-inflammatory, mucolytic, and antiviral activities. N-Acetylcysteine can be used in organoid culture media for non-small cell lung and colorectal cancer organoids, expansion media for mouse/human liver and pancreas 3D organoids and LWRN media for colonic 3D organoids from human biopsies. It is a component of S7 medium in the optimized protocol for generation of functional stem cell-derived islets (SC-islets). N-Acetylcysteine can be orally administered.
Scientific Data
Application of N-Acetylcysteine in Culture and Characterization of Adult Stem Cell-derived Lung Organoids. Adult stem cells isolated from human lung biopsy tissue were embedded and then cultured for 20-60 days in lung organoid expansion medium, which includes N-Acetylcysteine, Y-27632 dihydrochloride (Catalog # 1254), HEPES (Catalog # 3173), A 83-01 (Catalog # 2939), SB 202190 (Catalog # 1264), Nicotinamide (Catalog # 4106), along with other reagents used in the human lung organoid culture protocol. Lung organoids were able to differentiate and exhibit markers for various lung cell types. Lung organoids were stained with (A) anti-Cytokeratin 5 (KRT5) (green) and with anti-p63/TP73L (Catalog # AF1916; red) to visualize basal cells, (B) Anti-Podoplanin (PDPN) (Novus Biologicals, Catalog # NB600-1015; green) to visualize alveolar type I cells and anti- p63/TP73L (Catalog # AF1916; red) to visualize basal cells, (C, D) Anti-MUC5AC (Novus Biologicals, Catalog # NBP2-15196; green) to visualize Goblet cells and anti-SOX2 (Catalog # MAB2018; red). All samples were counterstained with DAPI (Catalog # 5748; blue).
Application of N-Acetylcysteine in Culture and Characterization of Adult Stem Cell-derived Liver Organoids. Adult stem cell-derived liver organoids were cultured using N-Acetylcysteine, Y-27632 dihydrochloride (Catalog # 1254), HEPES (Catalog # 3173), A 83-01 (Catalog # 2939), Forskolin (Catalog # 1099), along with other reagents used in the human liver organoid culture protocol. After 3 days, liver organoid expansion medium was added for 7-10 days, followed by liver organoid differentiation medium, which includes N-Acetylcysteine, HEPES (Catalog # 3173), Nicotinamide (Catalog # 4106), A 83-01 (Catalog # 2939), along with other reagents. Differentiated human liver organoids were stained using a (A) Human Serum Albumin Antibody (Catalog # MAB1455; red) and a (B) Human HNF-3beta /FoxA2 Antibody (Catalog # AF2400; red) and counterstained with DAPI (Catalog # 5748; blue).
Application of N-Acetylcysteine in Culture and Characterization of iPSC-derived Human Intestinal Organoids. iPSC-derived human intestinal organoids were cultured using N-Acetylcysteine, Y-27632 dihydrochloride (Catalog # 1254), SB 202190 (Catalog # 1264), A 83-01 (Catalog # 2939), Nicotinamide (Catalog # 4106), along with other reagents used in the human intestinal organoid culture protocol. (A) Human intestinal organoids were stained using a Human/Mouse/Rat Vimentin Antibody (Catalog # MAB2105; green) and a Human/Mouse Desmin Antibody (Catalog # AF3844; red) to visualize myofibroblast cells and counterstained with DAPI (Catalog # 5748; blue). (B) Human intestinal organoids were stained using a Human/Mouse E-Cadherin Antibody (Catalog # AF748; green), a Human MUC2 Antibody (Novus Biologicals, Catalog # NBP2-44431; red) and counterstained with DAPI (Catalog # 5748; blue).
Application of N-Acetylcysteine in Culture and Characterization of Adult Stem Cell-derived Human Descending Colon Organoids. Adult stem cells isolated from human descending colon were embedded and then cultured for 30 days in intestinal organoid culture medium, which includes N-Acetylcysteine, Y-27632 dihydrochloride (Catalog # 1254), SB 202190 (Catalog # 1264), A 83-01 (Catalog # 2939), Nicotinamide (Catalog # 4106), along with other reagents used in the human intestinal organoid culture protocol. (A) Organoids were fixed and stained with Anti-MUC2 (Novus Biologicals; Catalog # NBP2-44431; green) to visualize intestinal goblet cells and counterstained for E-Cadherin (Catalog # AF3844; red) and DAPI (Catalog # 5748; blue). (B) Organoids were fixed and stained with Human Chromogranin A Antibody(Catalog # MAB90981; green) to visualize enteroendocrine cells and counterstained for E-Cadherin (Catalog # AF3844; red) and DAPI (Catalog # 5748; blue).
Technical Data for N-Acetylcysteine
| M. Wt | 163.2 |
| Formula | C5H9NO3S |
| Storage | Store at -20°C |
| Purity | ≥98% (HPLC) |
| CAS Number | 616-91-1 |
| PubChem ID | 12035 |
| InChI Key | PWKSKIMOESPYIA-BYPYZUCNSA-N |
| Smiles | [C@@H](NC(C)=O)(C(O)=O)CS |
The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.
Tocris products are intended for laboratory research use only, unless stated otherwise.
Solubility Data for N-Acetylcysteine
| Solvent | Max Conc. mg/mL | Max Conc. mM | |
|---|---|---|---|
| Solubility | |||
| DMSO | 8.16 | 50 | |
| water | 8.16 | 50 |
Preparing Stock Solutions for N-Acetylcysteine
The following data is based on the product molecular weight 163.2. Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.
| Concentration / Solvent Volume / Mass | 1 mg | 5 mg | 10 mg |
|---|---|---|---|
| 0.5 mM | 12.25 mL | 61.27 mL | 122.55 mL |
| 2.5 mM | 2.45 mL | 12.25 mL | 24.51 mL |
| 5 mM | 1.23 mL | 6.13 mL | 12.25 mL |
| 25 mM | 0.25 mL | 1.23 mL | 2.45 mL |
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Product Datasheets for N-Acetylcysteine
References for N-Acetylcysteine
References are publications that support the biological activity of the product.
Cattaneo et al (2019) Tumor organoid-T-cell coculture systems. Nat.Protoc. 15 15 PMID: 31853056
Broutier et al (2016) Culture and establishment of self-renewing human and mouse adult liver and pancreas 3D organoids and their genetic manipulation. Nat.Protoc. 11 1724 PMID: 27560176
Balboa et al (2022) Functional, metabolic and transcriptional maturation of human pancreatic islets derived from stem cells. Nat.Biotechnol. 40 1042 PMID: 35241836
Moiseeva et al (2023) Senescence atlas reveals an aged-like inflamed niche that blunts muscle regeneration. Nature 613 169 PMID: 36544018
Geiler et al (2009) N-acetyl-L-cysteine (NAC) inhibits virus replication and expression of pro-inflammatory molecules in A549 cells infected with highly pathogenic H5N1 influenza A virus. Biochem.Pharmacol. 79 413 PMID: 19732754
If you know of a relevant reference for N-Acetylcysteine, please let us know.
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Citations for N-Acetylcysteine
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By
Chi Ngo on 08/18/2025
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: Gastric cancer cells (AGS)
NAC was used to inhibit ROS overproduction by Helicobacter pylori (H. pylori) injection in gastric cancer cells AGS. The use of ROS quencher NAC was to investigate the involvement of H.pylori-induced ROS overproduction in epithelial-mesenchymal transition of gastric cancer cells AGS.
NAC is water soluble. Made a 1000x stock solution and add it directly to the culture medium.
