| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
This compound serves as a model substrate for studying transglutaminase (tTG/TG2) activity and protein crosslinking. It is not a drug targeting a therapeutic receptor; rather, it is the molecular product of tTG-mediated crosslinking. tTG is the enzyme responsible for its formation, and the isopeptide bond plays a critical role in ECM stabilization and tissue remodeling. It is implicated in diseases including non-diabetic kidney disease, diabetic nephropathy, and glaucoma filtration.
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| ln Vitro |
In vitro, the epsilon-(gamma-glutamyl)-lysine isopeptide bond is used as a biomarker of tTG activity. Elevated levels of these crosslinks are observed in conditions with increased tTG expression or activity. The compound serves as a biochemical tool to study tTG-mediated protein crosslinking, which is crucial for stabilizing various biological structures including the extracellular matrix and fibrin clots. It can be used in enzyme activity assays to measure tTG activity by monitoring the formation of crosslinks using labeled substrates.
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| ln Vivo |
In vivo, epsilon-(gamma-glutamyl)-lysine crosslink levels are increased in disease states characterized by fibrosis, ECM remodeling, or enhanced tTG activity, including non-diabetic kidney disease, diabetic nephropathy, glaucoma, and certain fibrotic conditions. In transglutaminase-mediated reactions, these crosslinks contribute to tissue stabilization, wound healing, and neurodegenerative disease pathogenesis. Altered levels are studied as potential biomarkers of disease progression.
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| Enzyme Assay |
Standard tTG activity assay: coat 96-well microtiter plates with N,N-dimethylated casein (0.5-1 ug/well) in PBS overnight at 4degC. Block with 3% BSA in PBS for 1 hour at room temperature. Add tTG enzyme (0.1-1 ug/mL) in reaction buffer (50 mM Tris-HCl, 5 mM CaCl2, 10 mM DTT, pH 7.5) containing biotinylated pentylamine (50-200 uM) and varying concentrations of epsilon-(gamma-glutamyl)-lysine peptide (as competitor, 1 nM-10 uM). Incubate for 30-60 minutes at 37degC. Wash, add HRP-conjugated streptavidin, incubate for 30 minutes, wash again, add TMB substrate, measure absorbance at 450 nm. Alternatively, use fluorescently labeled substrate (e.g., monodansylcadaverine) and monitor fluorescence (excitation 360 nm, emission 550 nm).
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| Cell Assay |
For crosslink formation studies, culture fibroblasts or other ECM-producing cells in DMEM with 10% FBS. Induce tTG expression or activation with TGF-beta1 (2-10 ng/mL) for 24-72 hours. Lyse cells or collect ECM. Perform Western blot analysis using antibodies specific for epsilon-(gamma-glutamyl)-lysine crosslinks (available as monoclonal antibodies). Alternatively, use biotinylated substrates followed by streptavidin detection to visualize crosslinked proteins. For quantification, perform LC-MS/MS analysis after proteolytic digestion to measure the epsilon-(gamma-glutamyl)-lysine dipeptide in protein hydrolysates. For cell viability assays, treat cells with epsilon-(gamma-glutamyl)-lysine (10-500 uM) for 6-48 hours, assess toxicity by MTT or LDH assays.
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| Animal Protocol |
For disease model studies, use transglutaminase 2 (TG2, tTG) knockout mice or wild-type controls subjected to disease models such as unilateral ureteral obstruction (UUO) for kidney fibrosis, streptozotocin (STZ)-induced diabetic nephropathy, or tissue injury models. Administer TG2 inhibitors (e.g., cystamine, 10-50 mg/kg) to modulate crosslink formation. Collect tissue samples (kidney, liver, skin, eye) at sacrifice. Process for histology or homogenize for protein extraction. Quantify epsilon-(gamma-glutamyl)-lysine crosslink levels by ELISA or LC-MS/MS as a pharmacodynamic biomarker of tTG activity and ECM remodeling. For glaucoma models, use ocular hypertension models and measure crosslinks in the trabecular meshwork.
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| ADME/Pharmacokinetics |
Epsilon-(gamma-glutamyl)-lysine is an endogenous dipeptide that is rapidly catabolized or turned over as part of ECM remodeling. As a small hydrophilic peptide (MW approximately 275.3 for the free dipeptide; TFA salt MW 389.32), it is expected to have rapid renal clearance and a short plasma half-life (minutes) if introduced exogenously. Endogenous levels vary by tissue, typically in the picomole to nanomole per milligram protein range. Under normal homeostasis, crosslink formation is balanced by proteolytic ECM degradation.
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| Toxicity/Toxicokinetics |
Epsilon-(gamma-glutamyl)-lysine is an endogenous metabolite and is not considered toxic at physiological or elevated levels. However, excessive tTG-mediated crosslinking in fibrotic diseases contributes to pathological tissue stiffening and organ dysfunction. The compound itself does not have direct toxicity. Material safety data sheets note that under fire conditions, it may decompose and emit toxic fumes. Standard laboratory precautions (gloves, lab coat, safety glasses) should be used. The TFA salt may cause irritation to eyes, skin, and respiratory tract if handled improperly.
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| References |
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| Additional Infomation |
This compound is also known as H-Glu(H-Lys-OH)-OH TFA, gamma-Glu-ε-Lys TFA, or epsilon-(gamma-glutamyl)lysine. It serves as a model substrate or inhibitor in studies of transglutaminase activity and protein crosslinking, and is used in biochemical assays to investigate the role of epsilon-(gamma-glutamyl)-lysine linkages in tissue stabilization, wound healing, and neurodegenerative diseases. The TFA salt form enhances solubility in aqueous buffers. For research use only.
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| Molecular Formula |
C13H22F3N3O7
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| Molecular Weight |
389.32
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| Related CAS # |
Epsilon-(gamma-glutamyl)-lysine;17105-15-6
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| Appearance |
Typically exists as solid at room temperature
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO :~250 mg/mL (~642.15 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.34 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.08 mg/mL (5.34 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. 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. View More
Solubility in Formulation 3: ≥ 2.08 mg/mL (5.34 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5686 mL | 12.8429 mL | 25.6858 mL | |
| 5 mM | 0.5137 mL | 2.5686 mL | 5.1372 mL | |
| 10 mM | 0.2569 mL | 1.2843 mL | 2.5686 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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