Real Thiol
Based on 1 Customer Validation
Real Thiol is a reversible reaction-based fluorescent probe which can quantitatively monitor the real-time glutathione dynamics in living cells.
For research use only. We do not sell to patients.
- Purity: 98.37%
- CAS No.: 2280796-90-7
- Formula: C20H17N3O7
- Molecular Weight:411.36
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Storage:
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Protocol
- Purity & Documentation
- References
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Help & FAQs
Biological Activity
Real Thiol (RT) shows ratiometric fluorescence responses with a wide dynamic range when reacting with Glutathione (GSH). Real Thiol and its GSH adduct (RT-GSH) shows fluorescence maxima at 487 and 562 nm with excitation wavelengths at 405 and 488 nm, respectively. The Kd for the reaction between Real Thiol and GSH is 3.7 mM. Real Thiol has much improved quantum yields and photostability. The wide dynamic range of Real Thiol allows for monitoring of the GSH level changes within 1 to 10 mM in both directions in living cells. Gel permeation chromatography (GPC) analysis showes that 10% of Real Thiol reacts with thiolated proteins while 90% of Real Thiol reacts with GSH[1].
Guide (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)[1].
1. Dissolve Real Thiol in dimethyl sulfoxide (DMSO) to obtain a 10 mM stock solution, and perform further dilution before use.
2. Subsequently, resuspend the cells in fresh medium containing 1 μM Real Thiol, and stain for approximately 10 minutes.
3. Analyze using a flow cytometer.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Chemical Information
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CAS No. 2280796-90-7
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Appearance Solid
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Molecular Weight 411.36
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Formula C20H17N3O7
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Color Pink to red
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SMILES
O=C1OC2=CC(N3CCC3)=CC=C2C=C1/C=C(C(N(CC(O)=O)CC(O)=O)=O)\C#N
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
4°C, protect from light
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)
Solvent & Solubility
DMSO : ≥ 101 mg/mL (245.53 mM; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
* "≥" means soluble, but saturation unknown.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Protocol
Real Thiol (RT) stock solution is diluted with PBS to the desired concentrations. Equal volumes (typically 20 to 100 μL) of various Glutathione (GSH) solutions with different concentrations are mixed with the Real Thiol solution 10 to 15 min before measurement. All samples are prepared on one 96-well/384-well plate with 3 replicates each. The plate reader is set to read absorption of all samples first. Fluorescent signals are then recorded at λex=405 nm, λem=485 nm, and λex=488 nm, λem=565 nm with bottom read. For testing fluorescent interference by environmental factors, the Real Thiol stock solution is directly diluted with corresponding solutions (for example, certain pH buffer or glycerol solution) and measured using the plate reader with the same settings stated above[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Purity & Documentation
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Data Sheet (273 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
References
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 2.4310 mL | 12.1548 mL | 24.3096 mL | 60.7740 mL |
| 5 mM | 0.4862 mL | 2.4310 mL | 4.8619 mL | 12.1548 mL | |
| 10 mM | 0.2431 mL | 1.2155 mL | 2.4310 mL | 6.0774 mL | |
| 15 mM | 0.1621 mL | 0.8103 mL | 1.6206 mL | 4.0516 mL | |
| 20 mM | 0.1215 mL | 0.6077 mL | 1.2155 mL | 3.0387 mL | |
| 25 mM | 0.0972 mL | 0.4862 mL | 0.9724 mL | 2.4310 mL | |
| 30 mM | 0.0810 mL | 0.4052 mL | 0.8103 mL | 2.0258 mL | |
| 40 mM | 0.0608 mL | 0.3039 mL | 0.6077 mL | 1.5194 mL | |
| 50 mM | 0.0486 mL | 0.2431 mL | 0.4862 mL | 1.2155 mL | |
| 60 mM | 0.0405 mL | 0.2026 mL | 0.4052 mL | 1.0129 mL | |
| 80 mM | 0.0304 mL | 0.1519 mL | 0.3039 mL | 0.7597 mL | |
| 100 mM | 0.0243 mL | 0.1215 mL | 0.2431 mL | 0.6077 mL |
Real Thiol Related Classifications
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