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Real Thiol is a reversible reaction-based fluorescent probe which can quantitatively monitor the real-time glutathione dynamics in living cells.

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Real Thiol Chemical Structure

Real Thiol Chemical Structure

CAS No. : 2280796-90-7

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1 mg USD 350 In-stock
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Based on 1 publication(s) in Google Scholar

Other Forms of Real Thiol:

Real Thiol Related Antibodies

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Description

Real Thiol is a reversible reaction-based fluorescent probe which can quantitatively monitor the real-time glutathione dynamics in living cells.

In Vitro

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].
Guidelines (Following is our recommended protocol. This protocol only provides a guideline, and should be modified according to your specific needs)[1].
1. Real Thiol (RT) is dissolved in DMSO to obtain a 10 mM stock solutions and further diluted before use.
2. Cells are cultured in 10% FBS DMEM containing different concentrations of Buthionine sulfoximine (0.9, 1.9, 3.9, 7.8, 15.6, 31.3, 62.5, 125, 250 and 500?μM) for 72?h and harvested for analysis.
3. Cells are then suspended in fresh medium containing 1?μM of Real Thiol probe as single-cell suspensions, and keep the staining time between 9 and 10?min for each sample.
4. Analyze sample on a flow cytometer. PE and pacific-blue channels were selected to measure the fluorescence intensities of unreacted and reacted Real Thiol, respectively.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Real Thiol Related Antibodies
Molecular Weight

411.36

Formula

C20H17N3O7
CAS No.
Appearance

Solid

Color

Pink to red

SMILES

O=C1OC2=CC(N3CCC3)=CC=C2C=C1/C=C(C(N(CC(O)=O)CC(O)=O)=O)\C#N
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

Solvent & Solubility
In Vitro: 

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.

Preparing
Stock Solutions
Concentration Solvent Mass 1 mg 5 mg 10 mg
1 mM 2.4310 mL 12.1548 mL 24.3096 mL
5 mM 0.4862 mL 2.4310 mL 4.8619 mL
View the 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.

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Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

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Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

This equation is commonly abbreviated as: C1V1 = C2V2

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In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:

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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Calculation results:
Working solution concentration: mg/mL
Purity & Documentation

Purity: 99.87%

COA (249 KB) HNMR (233 KB) LCMS (112 KB)

Handling Instructions (2659 KB)
References
Kinase Assay
[1]

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].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.
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
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Real Thiol Related Classifications

Help & FAQs
  • Do most proteins show cross-species activity?

    Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

Keywords:

Real Thiol2280796-90-7Fluorescent DyeInhibitorinhibitorinhibit

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