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1mg |
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5mg |
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10mg |
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Other Sizes |
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ln Vitro |
TrxR-mediated disulfide bond breaking and subsequent intramolecular cyclization liberate the veiled naphthalimide fluorophore, causing TRFS-green to exhibit a green fluorescence switching change [1]. TRFS-green exhibits a faint signal at about 480 nm when stimulated at 377 nm. The creation of live cell-based TrxR inhibitor screening assays is aided by the pretreatment of living cells with TrxR inhibitors in a dose-dependent manner, which can block the fluorescence signal of TRFS-green in living cells [1]. The two main thiols in cells, GSH and Cys, as well as endogenous reducing agents like vitamin C and NADPH, have no effect on TRFS-green [1]. Assess TrxR activity in crude cell extracts [1]: Use a fluorometer (Em=540 nm, Ex=440 nm) to assess the fluorescence intensity after incubating cell lysates with TRFS-green for the recommended amount of time (about 1-2 hours). Moreover, the time-dependent increase in fluorescence over a certain duration (Em=540 nm, Ex=440 nm) can be observed. Through the microscope's green fluorescence channel, cells treated with 10 μM with TRFS-green for 2-4 hours can be seen [1]. Note: It is advised to wash the cells with PBS or new medium right before taking pictures in order to remove any remaining TRFS-green in order to achieve clean fluorescence images. The first fluorescent probe for TrxR with green emission is called TRFS-green. Nevertheless, TRFS-green reacts slowly to enzymes, and it takes more than two hours to produce the maximum fluorescence signal—even in the case of reducing agent tris(2-carboxyethyl)phosphine (TCEP) [2].
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References |
[1]. Liangwei Zhang, et al. Highly selective off-on fluorescent probe for imaging thioredoxin reductase in living cells. J Am Chem Soc. 2014 Jan 8;136(1):226-33.
[2]. Xinming Li, et al. A fast and specific fluorescent probe for thioredoxin reductase that works via disulphide bond cleavage. Nat Commun. 2019 Jun 21;10(1):2745. |
Molecular Formula |
C20H20N2O4S2
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Molecular Weight |
416.51
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CAS # |
1513848-14-0
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SMILES |
S1CC(CS1)OC(NC1=CC=C2C(N(C(C3C=CC=C1C2=3)=O)CCCC)=O)=O
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Solubility (In Vitro) |
DMSO: 25 mg/mL (60.02 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 | 2.4009 mL | 12.0045 mL | 24.0090 mL | |
5 mM | 0.4802 mL | 2.4009 mL | 4.8018 mL | |
10 mM | 0.2401 mL | 1.2005 mL | 2.4009 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.