| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
Intracellular glutathione (GSH) is the primary molecular target. The Real Thiol probe, generated intracellularly after esterase cleavage of the AM group, reversibly reacts with GSH to allow quantitative, real-time monitoring of GSH dynamics.
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|---|---|
| ln Vitro |
Acetoxymethyl (AM) esters, which are easily hydrolyzed by esterases to regenerate Real Thiol inside cells, are created from the carboxylic acid groups to increase the permeability of Real Thiol inside cells. When cells are treated with RT-AM, the fluorescence intensity values for the 405 nm channel are divided by the 488 nm channel at each corresponding pixel to create real-time ratiometric pictures of the rapid changes of intracellular GSH concentrations in single cells[1].
Real Thiol (the active form) is a reversible reaction-based fluorescent probe that quantitatively monitors the real-time glutathione dynamics in living cells. The fluorescence signal correlates with GSH concentration, and the reversibility allows tracking of both increases and decreases in GSH levels in response to stimuli. |
| ln Vivo |
There is no specific in vivo activity data available for this compound. RT-AM is designed for use in live cell imaging to monitor endogenous GSH fluctuations in real time. It is not intended for direct in vivo use in animals.
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| Enzyme Assay |
A standard protocol involves incubating RT-AM (e.g., 5-10 uM) with varying concentrations of glutathione (GSH, 0-10 mM) in a buffer solution (e.g., PBS, pH 7.4) at 37degC. The fluorescence signal is measured over time to characterize the reversibility and sensitivity of the probe. For control reactions, oxidizing agents (e.g., H2O2) can be added to observe signal changes.
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| Cell Assay |
A general protocol for cellular imaging involves seeding cells in a confocal dish and incubating with RT-AM (e.g., 5-10 uM) for 30 minutes at 37degC. The AM ester group facilitates cellular uptake, and once inside cells, esterases cleave the AM group to trap the active Real Thiol probe. After washing, live-cell imaging is performed. To induce dynamic changes in GSH, cells are treated with an oxidant (e.g., H2O2 or diamide) to decrease GSH or with a reducing agent (e.g., DTT) to increase GSH. Fluorescence changes are recorded in real time.
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| Animal Protocol |
A typical protocol for in vivo imaging would involve intravenous or intraperitoneal injection of RT-AM (e.g., 1-10 mg/kg) into a live animal model. After a suitable circulation time to allow for probe distribution and activation by esterases, the animal is placed under an in vivo imaging system to monitor GSH dynamics. However, this is a less common application for this probe.
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| ADME/Pharmacokinetics |
General PK properties for AM ester probes: The AM ester group enhances cell permeability, allowing entry into cells. Once inside, intracellular esterases cleave the AM group, trapping the active probe. Formulation can be in DMSO as a stock solution, with dilutions in PBS or culture medium.
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| Toxicity/Toxicokinetics |
General toxicity for imaging probes is considered low. No specific toxicity data is available for RT-AM. For similar AM ester probes, working concentrations up to 10 uM for 24 hours in cell models show good biocompatibility.
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| References | |
| Additional Infomation |
RT-AM is a research tool for studying redox biology, specifically the dynamic regulation of glutathione, the most abundant intracellular thiol and key antioxidant. Its reversible reaction mechanism is a significant advantage over irreversible GSH probes, allowing real-time monitoring of both GSH depletion and recovery. This product is for research use only.
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| Molecular Formula |
C26H25N3O11
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|---|---|
| Molecular Weight |
555.490207433701
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| Exact Mass |
555.148
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| CAS # |
2280796-94-1
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| Related CAS # |
Real Thiol;2280796-90-7
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| PubChem CID |
134128279
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| Appearance |
Pink to red solid powder
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| LogP |
1.8
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
13
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| Rotatable Bond Count |
15
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| Heavy Atom Count |
40
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| Complexity |
1110
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| Defined Atom Stereocenter Count |
0
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| SMILES |
O1C(C(/C=C(\C#N)/C(N(CC(=O)OCOC(C)=O)CC(=O)OCOC(C)=O)=O)=CC2=CC=C(C=C12)N1CCC1)=O
|
| InChi Key |
CLCCMBNDDZVBCA-AWQFTUOYSA-N
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| InChi Code |
InChI=1S/C26H25N3O11/c1-16(30)36-14-38-23(32)12-29(13-24(33)39-15-37-17(2)31)25(34)20(11-27)9-19-8-18-4-5-21(28-6-3-7-28)10-22(18)40-26(19)35/h4-5,8-10H,3,6-7,12-15H2,1-2H3/b20-9+
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| Chemical Name |
acetyloxymethyl 2-[[2-(acetyloxymethoxy)-2-oxoethyl]-[(E)-3-[7-(azetidin-1-yl)-2-oxochromen-3-yl]-2-cyanoprop-2-enoyl]amino]acetate
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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 |
| 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: 25 mg/mL (45.01 mM)
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|---|---|
| Solubility (In Vivo) |
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.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *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). View More
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)] Oral Formulations
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). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.8002 mL | 9.0011 mL | 18.0021 mL | |
| 5 mM | 0.3600 mL | 1.8002 mL | 3.6004 mL | |
| 10 mM | 0.1800 mL | 0.9001 mL | 1.8002 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.
Products are for research use only; We do not sell to patients
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