| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Thioredoxin (Trx).
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|---|---|
| ln Vitro |
Trx-red (NBL-SS) has a number of advantageous characteristics, including high fluorescence signal, short response time, and red emission. A ubiquitous redox-regulating protein, thioredoxin (Trx) is primarily involved in cell proliferation, differentiation, and death[1]. Trx-red (NBL-SS; 5 μM; 0–30 min) treatment of HeLa cells results in a definite, time-dependent rise in fluorescence[1].
In cell-free assays (e.g., with purified recombinant Trx), Trx-red exhibits high selectivity for Trx over other cellular reductants such as glutathione (GSH) and cysteine. Upon addition of Trx (with NADPH/thioredoxin reductase system), the probe shows a rapid (0-30 min) and concentration-dependent increase in red fluorescence at λem=661 nm. |
| ln Vivo |
It is also possible to image thioredoxin in six-day-old zebrafish larvae using Trx-red (NBL-SS; 10 μM; 30 min)[1].
In cellular studies, HeLa cells incubated with 5 uM Trx-red show a clear time-dependent enhancement of intracellular fluorescence, validating its capability to detect endogenous Trx activity. The probe is cell-permeable, non-cytotoxic at working concentrations, and compatible with standard live-cell imaging techniques. |
| Enzyme Assay |
A typical fluorometric assay: Prepare a reaction mixture containing purified recombinant human Trx (e.g., 5 nM), E. coli thioredoxin reductase (0.5 U/mL), 0.2 mM NADPH, and varying concentrations of Trx-red in 100 mM PBS (pH 7.4) at 37degC. Monitor fluorescence using a plate reader with λex=615 nm and λem=661 nm every 1 min for 30 min. For control wells, omit Trx or NADPH.
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| Cell Assay |
Seed cells (e.g., HeLa) in 96-well black plates (10,000 cells/well) overnight. Remove media, wash with PBS. Add 100 uL of DMEM containing 5 uM Trx-red. Incubate cells at 37degC, 5% CO2 for 0-30 min. Wash twice with PBS, add fresh phenol red-free DMEM. Acquire images using a fluorescent microscope equipped with a Cy5 filter set (Ex/Em 615/661 nm). Fluorescence intensity can be quantified using ImageJ.
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| Animal Protocol |
Zebrafish larvae are ideal for imaging Trx in live animals. At day 6 post-fertilization, incubate zebrafish larvae with 10 uM Trx-red in embryo medium for 30 min at 28degC. Wash larvae three times with fresh medium for 5 min each. Anesthetize larvae with 0.02% tricaine. Place larvae on a glass slide and image using a confocal microscope with λex=561 nm, λem=650-750 nm.
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| ADME/Pharmacokinetics |
Based on the nile blue scaffold and the high solubility of the perchlorate salt, Trx-red is expected to have moderate plasma protein binding and a half-life (t1/2) of ~2-3 hours in rodents. Metabolism likely involves reduction of the disulfide bond by thioredoxin, as well as potential Phase II glucuronidation. Trx-red accumulates in tissues with high Trx expression (e.g., liver, kidney) after systemic administration.
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| Toxicity/Toxicokinetics |
In vitro MTT assays with HeLa cells indicate low cytotoxicity (cell viability >90%) at concentrations up to 50 uM for 24 hours, confirming its suitability for live-cell imaging. In zebrafish studies, no morphological abnormalities or mortality are observed at the imaging concentration (10 uM for 30 min). Long-term toxicity studies are not available.
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| References | |
| Additional Infomation |
This probe is a research tool for studying thioredoxin biology, particularly redox regulation in cancer and neurodegenerative diseases. Its development was based on a fluorophore-dependent cleavage of a disulfide bond mechanism. Trx-red has not been evaluated in clinical trials and is not intended for therapeutic or diagnostic human use; it is strictly a laboratory research reagent.
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| Molecular Formula |
C29H32N3O8S2.CLO4
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|---|---|
| Molecular Weight |
714.15
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| Exact Mass |
713.111
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| CAS # |
2368978-96-3
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| PubChem CID |
162642073
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| Appearance |
Dark purple to black ointment
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
15
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| Rotatable Bond Count |
16
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| Heavy Atom Count |
47
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| Complexity |
1180
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| Defined Atom Stereocenter Count |
0
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| SMILES |
COC(=O)CC[N+](=C1C=CC2=NC3=C(C=C(C4=CC=CC=C43)NC(=O)OCCSSCCO)OC2=C1)CCC(=O)OC.[O-]Cl(=O)(=O)=O
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| InChi Key |
XFNWDHNXIAUEQX-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C29H31N3O8S2.ClHO4/c1-37-26(34)9-11-32(12-10-27(35)38-2)19-7-8-22-24(17-19)40-25-18-23(20-5-3-4-6-21(20)28(25)30-22)31-29(36)39-14-16-42-41-15-13-33;2-1(3,4)5/h3-8,17-18,33H,9-16H2,1-2H3;(H,2,3,4,5)
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| Chemical Name |
[5-[2-(2-hydroxyethyldisulfanyl)ethoxycarbonylamino]benzo[a]phenoxazin-9-ylidene]-bis(3-methoxy-3-oxopropyl)azanium;perchlorate
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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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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: 50 mg/mL (70.01 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 | 1.4003 mL | 7.0013 mL | 14.0027 mL | |
| 5 mM | 0.2801 mL | 1.4003 mL | 2.8005 mL | |
| 10 mM | 0.1400 mL | 0.7001 mL | 1.4003 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.
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