Diamide (Synonyms: 1,1′-Azobis(N,N-dimethylformamide), N,N,N′,N′-Tetramethylazodicarboxamide, Azodicarboxylic acid bis(dimethylamide), Diazenedicarboxylic acid bis(N,N-dimethylamide), TMAD；) |
Catalog No.GD10507
One-Click Copy Product Info
|
Diamide is a commonly used thiol oxidant, also recognized as a highly cell-permeable reagent that can oxidize reduced glutathione (GSH) to oxidized glutathione (GSSG) without generating reactive oxygen species (ROS).
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 10465-78-8
Sample solution is provided at 25 µL, 10mM.
_1-3.$$$39978408@51.jpg');)
_1-9.$$$38538795@65.jpg');)
_1-9.$$$39112701@51.jpg');)
_1-7.$$$37316672@70.jpg');)
_366-372.$$$36198801@70.jpg');)
.$$$38565142@65.jpg');)
_1867-1883.$$$33248023@67.jpg');)
_eads6055.$$$39977546@45.jpg');)
_eadm9805.$$$40080571@45.jpg');)
_eadj3020.$$$39666821@45.jpg');)
_1-20.$$$39757301@28.jpg');)
_1-24.$$$38898113@28.jpg');)
_273-287.$$$36806353@46.jpg');)
_1-16.$$$37156877@44.jpg');)
_1-19.$$$37460805@44.jpg');)
_1291-1307.$$$34518654@46.jpg');)
Diamide is a commonly used thiol oxidant, also recognized as a highly cell-permeable reagent that can oxidize reduced glutathione (GSH) to oxidized glutathione (GSSG) without generating reactive oxygen species (ROS)[1-2]. Diamide has been widely applied in cell-free systems, in vitro cellular experiments, and in vivo studies to rapidly oxidize GSH and/or protein thiols[3-4].
In vitro, Diamide (0–400μM) treatment of LoVo cells significantly reversed the increase in intracellular lactate levels induced by DDR1a knockdown and counteracted the regulatory effect of DDR1a on pyruvate kinase activity[5]. Diamide (0.1–0.5mM) treatment of rat aortic smooth muscle cells concentration-dependently increased protein glutathionylation, induced mitochondrial proton leak, and reduced ATP-linked oxygen consumption; higher concentrations (≥0.25mM) led to bioenergetic failure and cell death[6].
In vivo, stereotaxic injection of Diamide (1.45μmol) into the substantia nigra of C57BL/6 mice (single injection), Diamide significantly reduced glutathione levels in brain tissue, increased oxidative stress markers, and induced degeneration of dopaminergic neurons in the substantia nigra, which can be utilized to establish a mouse model of Parkinson's disease[7].
References:
[1] Kosower NS, Kosower EM, Koppel RL. Sensitivity of hemoglobin thiol groups within red blood cells of rat during oxidation of glutathione. Eur J Biochem. 1977 Aug 1;77(3):529-34.
[2] Kosower NS, Kosower EM, Wertheim B, et al. Diamide, a new reagent for the intracellular oxidation of glutathione to the disulfide. Biochem Biophys Res Commun. 1969 Nov 6;37(4):593-6.
[3] Liu Y, Min W. Thioredoxin promotes ASK1 ubiquitination and degradation to inhibit ASK1-mediated apoptosis in a redox activity-independent manner. Circ Res. 2002 Jun 28;90(12):1259-66.
[4] Marey-Semper I, Gelman M, Lévi-Strauss M. A selective toxicity toward cultured mesencephalic dopaminergic neurons is induced by the synergistic effects of energetic metabolism impairment and NMDA receptor activation. J Neurosci. 1995 Sep;15(9):5912-8.
[5] Xiong B, Xie Z, Song F, et al. DDR1 promotes LoVo cell proliferation by regulating energy metabolism. Acta Biochim Biophys Sin (Shanghai). 2022 May 25;54(5):615-624.
[6] Hill BG, Higdon AN, Dranka BP, et al. Regulation of vascular smooth muscle cell bioenergetic function by protein glutathiolation. Biochim Biophys Acta. 2010 Feb;1797(2):285-95.
[7] Ray A, Kambali M, Ravindranath V. Thiol Oxidation by Diamide Leads to Dopaminergic Degeneration and Parkinsonism Phenotype in Mice: A Model for Parkinson's Disease. Antioxid Redox Signal. 2016 Aug 10;25(5):252-67.
| Cell experiment [1]: | |
|
Cell lines |
LoVo cells (human colorectal carcinoma cell line) |
|
Preparation Method |
LoVo cells were maintained in ATCC-formulated Ham's F12K medium supplemented with 10% fetal bovine serum (FBS), 100U/mL penicillin, and 100μg/mL streptomycin at 37°C, 5% CO₂. LoVo cells were treated with diamide at concentrations of 0, 50, 100, 200, and 400μM for 15 minutes. |
|
Reaction Conditions |
0-400μM; 15h |
|
Applications |
Diamide significantly reversed DDR1a knockdown-induced intracellular lactate accumulation in LoVo cells by inhibiting pyruvate kinase (PK) activity. Diamide treatment also reduced phosphorylated AKT (p-AKT) levels and suppressed PI3K/AKT/PKM2 signaling pathway activation, thereby regulating glycolytic flux and metabolic reprogramming in CRC cells. |
| Animal experiment [2]: | |
|
Animal models |
C57BL/6 mice |
|
Preparation Method |
Mice were stereotaxically administered a single unilateral dose of Diamide (1.45μmol) into the substantia nigra (SN). Mice were sacrificed 14 days post-injection for behavioral, histopathological, and biochemical analyses. |
|
Dosage form |
1.45μmol; stereotaxic injection; Single administration. |
|
Applications |
Diamide injection led to hemiparkinsonian motor deficits, including reduced rotarod latency, impaired grip strength, and asymmetric contralateral rotations. Diamide selectively induced dopaminergic neurodegeneration in the ipsilateral SNpc and striatal fiber loss, accompanied by α-synuclein aggregation, glutathione depletion, and activation of the ASK1-p38 MAPK cell death signaling pathway. |
|
References: |
|
| Cas No. | 10465-78-8 | SDF | |
| Synonyms | 1,1′-Azobis(N,N-dimethylformamide), N,N,N′,N′-Tetramethylazodicarboxamide, Azodicarboxylic acid bis(dimethylamide), Diazenedicarboxylic acid bis(N,N-dimethylamide), TMAD； | ||
| Formula | C6H12N4O2 | M.Wt | 172.19 |
| Solubility | DMSO：>22mg/ml, water: 19.60-21.00 mg/mL, clear to slightly hazy, orange | Storage | Storage at -20 ° C, filled with argon |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 5.8075 mL | 29.0377 mL | 58.0754 mL |
| 5 mM | 1.1615 mL | 5.8075 mL | 11.6151 mL |
| 10 mM | 580.8 μL | 2.9038 mL | 5.8075 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid 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 master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >99.50% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 30 reference(s) in Google Scholar.)