Ammonium γ-glutamylcysteine (0-80 μM; 24 hours) inhibits M1 polarization of Raw264.7 macrophages by suppressing the JAK1/STAT1/3, AKT, MAPK and NF-κB signaling pathways and activating the AMPK/SIRT1 axis [1]. Ammonium γ-glutamylcysteine (0-80 μM; 24 hours) promotes M2 polarization of Raw264.7 macrophages and drives their repolarization from M1 to M2 by activating the IL-4/STAT6 and AMPK/SIRT1 signaling pathways [1]. Ammonium γ-glutamylcysteine (20-80 μM; 24 hours) induces Th2 polarization of human Jurcat CD4+ T lymphocytes by promoting IL-4 secretion [1]. Ammonium γ-glutamylcysteine (2 mM; 6 h) activates the Nrf2 signaling pathway via nuclear translocation and upregulates the expression of antioxidant target genes and proteins in primary intestinal epithelial cells (IECs) of chicken embryos [2]. Ammonium γ-glutamylcysteine (2 mM; 6 h) inhibits oxidative stress induced by Salmonella typhimurium in primary intestinal epithelial cells (IECs) of wild-type chicken embryos by maintaining the activated state of Nrf2 [2]. Ammonium γ-glutamylcysteine (2 mM; 6 h) protects primary intestinal epithelial cells (IECs) of wild-type chicken embryos from intestinal barrier disruption and inflammatory damage caused by Salmonella typhimurium, and this protective effect depends on the Nrf2 signaling pathway [2]. Ammonium γ-glutamylcysteine (20-80 μM; 24 hours after insulin + palmitate pretreatment) dose-dependently activated the IGF-1R/IRS1/PI3K/Akt signaling pathway in insulin-resistant HepG2 cells, primary mouse hepatocytes, and insulin + palmitate (PA)-induced C2C12 myotubes by increasing the phosphorylation level of key pathway proteins [3]. Ammonium γ-glutamylcysteine (2-4 mM; 2 hours after pretreatment, 12 hours after cadmium exposure) inhibited the activation of the JNK/p38 MAPK and PI3K/Akt signaling pathways in cadmium-treated cells by downregulating the expression of pro-apoptotic markers and normalizing the Bax/Bcl-2 ratio. Therefore, ammonium γ-glutamylcysteine can inhibit changes in cadmium-induced apoptosis-related proteins, inhibit cadmium-induced apoptosis, and prevent cadmium-induced depolarization of mitochondrial transmembrane potential in PC12 cells [4]. Ammonium γ-glutamylcysteine (2-4 mM; 2 hours after pretreatment followed by 12 hours of exposure to cadmium) inhibited cadmium-induced oxidative stress in PC12 cells in a dose-dependent manner by reducing reactive oxygen species (ROS) and lipid peroxidation levels, restoring antioxidant enzyme activity, and maintaining intracellular glutathione (GSH) homeostasis [4]. Ammonium γ-glutamylcysteine (0.25-16 mM; 24 hours) was non-toxic to BV-2 cells at concentrations up to 4 mM, while higher concentrations (8, 16 mM) reduced cell viability in a dose-dependent manner [5]. Ammonium γ-glutamylcysteine (2-4 mM; pretreatment for 30 min followed by AβO exposure for 24 h) dose-dependently inhibited the release of AβO-induced pro-inflammatory mediators (TNF-α, IL-1β, NO) and the expression of pro-inflammatory proteins (iNOS, COX-2) in BV-2 cells, inhibited AβO-induced oxidative stress, and restored the antioxidant capacity of cells [5]. Ammonium γ-glutamylcysteine (4 mM; pretreatment for 30 min followed by AβO exposure for 6-24 h) inhibited the activation of the AβO-induced NF-κB signaling pathway in BV-2 cells, upregulated and maintained the expression of Nurr1 mRNA and protein, thereby exerting an anti-inflammatory effect by inhibiting the binding of NF-κB p65 to the AβO-induced iNOS promoter in BV-2 cells [5]. Ammonium γ-glutamylcysteine (2-4 mM; pretreatment for 30 min followed by exposure to AβO for 24 h) dose-dependently inhibited the release of AβO-induced pro-inflammatory mediators (TNF-α, IL-1β, NO) in primary mouse microglia, suppressed oxidative stress, and restored the antioxidant capacity of cells [5]. Ammonium γ-glutamylcysteine (0.25-2 mM; 12 h) increased GSH levels, GSH/GSSG ratio, GPX activity, and cell viability in primary cortical neurons treated with OGD/R, with 2 mM treatment for 12 h showing the strongest effect [6]. Ammonium γ-glutamylcysteine (0.85-7 mM; 12 h) increased GSH levels, GSH/GSSG ratio, GPX activity, and cell viability in PC12 cells treated with oxygen-glucose deprivation/reoxygenation (OGD/R) [6]. Ammonium γ-glutamylcysteine (1.7-7 mM; 0-12 h, time course 0-12 h) can regulate the mRNA and protein levels of GSS in OGD/R treated PC12 cells, promote the nuclear translocation of Nrf2 in cells, reduce the interaction between Nrf2 and Keap1, inhibit the upregulation of Keap1, reduce MDA and Fe2+ levels, and improve cell viability, thereby inhibiting ferroptosis [6]. Ammonium γ-glutamylcysteine (3.5 mM; 12 h, time course 0-12 h) activates Nrf2 in OGD/R treated PC12 cells by increasing the level of phosphorylated PKC-ε [6]. Ammonium γ-glutamylcysteine (200 μM; pre-incubated for 15 min, then co-incubated with oligomer Aβ40 for 24 h) can protect primary human astrocytes from oligomer Aβ40-induced cytotoxicity, apoptosis, oxidative stress, neuroinflammation and dysregulation of metalloproteinase activity, while restoring antioxidant status and GSH levels [7]. Ammonium γ-glutamylcysteine (20-80 μM; pre-treated for 2 hours followed by 24 h of ethanol exposure; 400 μM; single 24 h treatment) protects human L02 hepatocytes from ethanol-induced damage by dose-dependently increasing cell viability, reducing liver enzyme release, inhibiting apoptosis, inhibiting oxidative stress and mitochondrial damage, and attenuating the activation of pro-inflammatory signaling pathways [9].

Ammonium γ-glutamylcysteine (600-1200 mg/kg; orally; daily; for about 3 days) significantly improved the survival rate of mice with TNBS-induced inflammatory bowel disease, reduced colonic damage, and converted macrophage polarization from a pro-inflammatory M1 phenotype to an anti-inflammatory M2 phenotype, with both doses showing therapeutic effects [1]. Ammonium γ-glutamylcysteine (250-500 mg/kg; orally; daily; for 8 weeks) dose-dependently improved glycemic control, insulin sensitivity, β-cell function, and hepatic steatosis in db/db mice, while reducing diabetes-related organ damage [3]. Ammonium γ-glutamylcysteine (100-400 mg/kg/d; orally; daily; for 20 days) dose-dependently inhibited AβO-induced neuroinflammation in male ICR mice [5]. Ammonium γ-glutamylcysteine (688 mg/kg; orally; single dose) upregulated GSH levels by activating the PKC-ε/Nrf2/GSS pathway and significantly reduced cerebral infarction volume, neurological dysfunction, and neuronal ferroptosis induced by cerebral ischemia/reperfusion injury in male Sprague-Dawley rats [6]. Ammonium γ-glutamylcysteine (600 mg/kg; orally; single dose) significantly reduced ds-HMGB1 levels in the dorsal root ganglia (DRG) of OIPN mice and reversed oxaliplatin-induced mechanical hyperalgesia [8]. Ammonium γ-glutamylcysteine (700-1200 mg/kg; orally; daily; 7 days) alleviated acute ethanol-induced hepatotoxicity in male C57BL/6JNifdc mice in a dose-dependent manner by reducing hepatic enzyme release, restoring hepatic antioxidant levels, alleviating histopathological damage, and inhibiting inflammatory signaling pathways [9].

Western Blot Analysis [3]
Cell Types:Insulin + palmitic acid induced insulin-resistant HepG2 cells, primary mouse hepatocytes, C2C12 mouse skeletal muscle myotubes
Test concentrations:0, 20, 40, 80 μM
Incubation Duration:24 hours (after 24 hours of insulin + PA pretreatment)
Experimental Results:Compared with cells treated with insulin + PA alone, the phosphorylation levels of p-IGF-1R (Tyr1135), p-IRS1 (Tyr612), p-PI3K and p-Akt (Ser473) in all three cell types showed a significant dose-dependent increase, with significant increases observed at 20, 40 and 80 μM.

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Apoptosis analysis [4]
Cell Types: PC12 cells
Tested Concentrations: 4 mM
Incubation Duration: 2 hours of pretreatment + 12 hours of cadmium exposure
Experimental Results: Significantly reduced the number of TUNEL-positive PC12 cells. Reduced the percentage of cadmium-induced apoptosis.

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Western Blot Analysis [4]
Cell Types: PC12 cells
Tested Concentrations: 4 mM
Incubation Duration: 2 hours of pretreatment + 12 hours of cadmium exposure
Experimental Results: The Bax/Bcl-2 ratio was downregulated in cadmium-treated PC12 cells. The protein levels of cytochrome c, cleaved-caspase-9, cleaved-caspase-3, and cleaved-PARP in cadmium-treated PC12 cells were reduced.

Animal/Disease Models:BALB/c (8-week-old males, 20-22 g, TNBS-induced colitis) [1]
Doses: 600 mg/kg; 1200 mg/kg
Route of Administration: Oral; once daily; approximately 3 days
Experimental Results: Significantly reduced mortality. Reversed TNBS-induced weight loss. Reduced the increase in Disease Activity Index (DAI). Alleviated colonic shortening, bleeding, and ulceration. Reduced histological damage score. Significantly reduced the mRNA levels of M1 markers (Inos, IL-1β) in colonic tissue. Significantly increased the mRNA levels of M2 markers (Cd206, Arg1) in colonic tissue. Serum TNF-α levels were significantly reduced compared to the TNBS model group. Western blotting and immunohistochemistry confirmed that the protein levels of M1 macrophage markers (iNOS, IL-1β) were decreased and the protein levels of M2 macrophage markers (CD206, ARG1) were increased in colonic tissue. TNBS-induced phosphorylation levels of JAK1, STAT1, STAT3, AKT, JNK, ERK, p38, IKKα/β, and IkBα were significantly decreased in colonic lamina propria mononuclear cells. STAT6 phosphorylation levels and serum IL-4 levels were increased.

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Animal/Disease Models: C57BL/6J mice (male, 6 weeks old, 20-22 g); db/db mice (male, 6 weeks old, 30-35 g, spontaneous leptin receptor mutation model) [3]
Doses: 250 mg/kg; 500 mg/kg
Route of Administration: Oral; once daily; for 8 weeks
Experimental Results: Food intake, food utilization and water consumption were significantly reduced in the 500 mg/kg dose group. Water consumption was reduced in the 250 mg/kg dose group. Fasting blood glucose, serum glycated hemoglobin (HbA1c), serum insulin level and homeostatic model insulin resistance index (HOMA-IR) of db/db mice decreased in a dose-dependent manner, while HOMA-β index increased. In db/db mice, dose-dependent improvement was observed in glucose tolerance (decreased area under the curve of the oral glucose tolerance test) and insulin sensitivity (decreased area under the curve of the insulin tolerance test). In db/db mice, dose-dependent improvement was observed in organ coefficients (decreased liver coefficient, increased heart and kidney coefficients) and reduced tissue damage (cardiomyocyte degeneration, glomerular/tubular injury, hepatocyte ballooning degeneration, islet loss), with the 500 mg/kg dose group showing superior efficacy compared to metformin. In db/db mice, dose-dependent reduction was observed in the urinary microalbumin/albumin ratio and increased liver glycogen content. In db/db mice, dose-dependent reduction was observed in the weight of subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) and reduced adipocyte volume. In db/db mice, the drug dose-dependently reduced hepatic triglyceride (TG), total cholesterol (TC), and serum TG, TC, and low-density lipoprotein cholesterol (LDL-C) levels, while increasing serum high-density lipoprotein cholesterol (HDL-C) levels; the 500 mg/kg dose group was more effective than metformin in reducing hepatic TG. The drug dose-dependently reduced the accumulation of lipid droplets in the liver of db/db mice (Oil Red O staining) and decreased serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels. The drug dose-dependently reduced the expression of CD36, PPARα, and CPT1A proteins in the liver of db/db mice and increased the expression of phosphorylated insulin-like growth factor-1 receptor (p-IGF-1R), phosphorylated insulin-like growth factor receptor substrate 1 (p-IRS1), phosphorylated PI3K, and phosphorylated Akt proteins in the liver. In C57BL/6J mice, no significant effects on body weight, fasting blood glucose, organ coefficient, or histology were observed at either dose.

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Animal/Disease Models:ICR mice (male, 22-25 g, intraventricular injection of 2 μg AβO to induce Alzheimer's disease model) [5]
Doses: 100 mg/kg/d; 400 mg/kg/d
Route of Administration: Oral; daily; 20 days
Experimental Results: Inhibited AβO-induced microglial activation (reduced Iba1 fluorescence intensity in hippocampus) and inhibited the upregulation of COX-2, iNOS and Iba1 protein expression in hippocampus (100 mg/kg/d dose). Compared with the 100 mg/kg/d dose, the 400 mg/kg/d dose more effectively inhibited AβO-induced microglial activation and significantly reduced Iba1 fluorescence intensity; compared with the 100 mg/kg/d dose, the 400 mg/kg/d dose more effectively inhibited the AβO-induced upregulation of hippocampal COX-2, iNOS and Iba1 protein expression.

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Animal/Disease Models:Sprague-Dawley (SD) mice (male, 260-300 g, induced transient focal cerebral ischemia by reperfusion after 90-minute middle cerebral artery occlusion) [6]
Doses: 688 mg/kg
Route of Administration: Oral; single dose (administered 1.5 hours after middle cerebral artery occlusion)
Experimental Results: Significantly reduced infarct volume. Significantly reduced neurological deficit scores. Significantly increased the number of Nissl staining positive neurons in the ipsilateral cerebral cortex. Significantly reduced the number of FJB+/NeuN+ (apoptotic) neurons in the cerebral cortex. Alleviated MCAO/R-induced neuronal mitochondrial damage, including reduced mitochondrial cristae loss and outer membrane rupture. Significantly reduced cortical H₂O₂, malondialdehyde (MDA) and Fe²⁺ levels and reduced the number of 4-HNE positive neurons. It restored MCAO/R-induced changes in ferroptosis-related protein and mRNA levels: increased FTH1 and GPX4 levels, and decreased ACSL4 and TF levels (with no effect on SLC7A11). It significantly increased cortical GSH levels, the GSH/GSSG ratio, and glutathione peroxidase (GPX) activity. It inhibited MCAO/R-induced decreases in cortical GSS mRNA and protein levels and increased the number of GSS/NeuN positive neurons. Total Nrf2 and phosphorylated Nrf2 protein levels were increased in cortical tissue, as were the number of Nrf2/NeuN and p-Nrf2/NeuN positive neurons. Phosphorylated PKC protein levels were also increased in cerebral cortex tissue.

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Animal/Disease Models:C57BL/6J (male, 8-10 weeks old, 22-26 g, SPF grade, oxaliplatin-induced peripheral neuropathy model) [8]
Doses: 600 mg/kg
Route of Administration: Oral; once a week 2 hours before oxaliplatin injection
Experimental Results: ds-HMGB1 levels in the dorsal root ganglion (DRG) decreased to about 2.0 relative units. Serum ds-HMGB1 levels decreased less, to about 1.6 relative units. The claw withdrawal threshold increased to about 1.2 g, reversing oxaliplatin-induced mechanical hyperalgesia. Cold avoidance behavior scores decreased to about 4.0, slightly improving cold sensitivity. Compared with equimolar glutathione, this product showed greater efficacy in reducing DRG ds-HMGB1 levels and improving pain response.

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Animal/Disease Models:C57BL/6JNifdc (male, 6-8 weeks old, acute ethanol-induced hepatotoxicity model) [9]
Doses: 700 mg/kg; 1200 mg/kg
Route of Administration: Oral; once daily; for 7 days
Experimental Results: Reduced ethanol-induced elevations in serum ALT, AST, and TG levels in a dose-dependent manner. Reversed ethanol-induced reductions in hepatic GSH levels. Reduced hepatic lipid droplet formation and inflammatory cell infiltration. Reduced ethanol-induced elevations in hepatic iNOS, p-p65, p-IKKα/β, and p-IκBα proteins in a dose-dependent manner.

[1]. γ-Glutamylcysteine rescues mice from TNBS-driven inflammatory bowel disease through regulating macrophages polarization. Inflamm Res. 2023 Mar;72(3):603-621.

[2]. Lactobacillus crispatus 7-4 Mitigates Salmonella typhimurium-Induced Enteritis via the γ‑Glutamylcysteine-Mediated Nrf2 Pathway. Probiotics Antimicrob Proteins. 2025;17(5):3378-3391.

[3]. γ-glutamylcysteine alleviates insulin resistance and hepatic steatosis by regulating adenylate cyclase and IGF-1R/IRS1/PI3K/Akt signaling pathways. J Nutr Biochem. 2023 Sep;119:109404.

[4]. γ-glutamylcysteine suppresses cadmium-induced apoptosis in PC12 cells via regulating oxidative stress. Toxicology. 2022;465:153029.

[5]. γ-Glutamylcysteine attenuates amyloid-β oligomers-induced neuroinflammation in microglia via blocking NF-κB signaling pathway. Chem Biol Interact. 2022;363:110019.

[6]. γ-Glutamylcysteine Exerts Neuroprotection Effects against Cerebral Ischemia/Reperfusion Injury through Inhibiting Lipid Peroxidation and Ferroptosis. Antioxidants (Basel). 2022;11(9):1653. Published 2022 Aug 25.

[7]. The Precursor to Glutathione (GSH), γ-Glutamylcysteine (GGC), Can Ameliorate Oxidative Damage and Neuroinflammation Induced by Aβ40 Oligomers in Human Astrocytes. Front Aging Neurosci. 2019;11:177. Published 2019 Aug 8.

[8]. Microglial macrophage-derived ds-HMGB1 in DRG orchestrates neuropathic pain through immune-neural signaling. Cell Rep. 2025;44(12):116671.

[9]. γ-Glutamylcysteine alleviates ethanol-induced hepatotoxicity via suppressing oxidative stress, apoptosis, and inflammation. J Food Biochem. 2022;46(10):e14318.

C8H17N3O5S

267.30

Typically exists as solids at room temperature

OC([C@H](CCC(N[C@H](C(O)=O)CS)=O)N)=O.N

γ-Glu-Cys ammonium

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)