| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
Thiol groups (-SH) in proteins and other biomolecules are the primary molecular targets. The probe is designed to specifically react with thiols, enabling their detection and labeling.
|
|---|---|
| ln Vitro |
Using Naph-EA-mal (Thiol-green 1) (1 μM; 5 mins) on Hep G2 cells, it is possible to image thiols in living cells [1]. A strong green fluorescence is seen. Less fluorescence is seen in cells when they are preincubated with NEM (a thiol block reagent; 100 μM) to inhibit the intracellular thiols[1]. Monitoring thiol-disulfide levels in fixed cells Naph-EA-mal (Thiol-green 1) (10 μM) causes high fluorescence in Hep G2 cells because most cellular thiols are present in the reduced form. Sharp dimming of the fluorescence signal is seen following diamide treatment[1]. After diamide (200 μM) oxidized the Hep G2 cells for 30 minutes, 70% EtOH fixed them, and NEM (100 μM) inhibited the residual free thiols. Subsequently, the cells undergo further treatment with TCEP (5 mM), and Naph-EA-mal (Thiol-green 1) (10 μM) is added. Once more, the green fluorescent signal is observed[1].
Naph-EA-mal can be used to detect thiols in living cells, label protein thiols, quantify the concentration of total thiols in cell lysate, and determine reversible protein thiol oxidation in fixed cells. It has an excitation wavelength of 488 nm and an emission wavelength of 540 nm. |
| ln Vivo |
There is no specific in vivo activity data available for this compound. However, as a fluorescent probe for thiols, similar compounds are typically used for real-time imaging of thiol-containing biomolecules in live cells and small animal models.
|
| Enzyme Assay |
A standard protocol involves incubating Naph-EA-mal (e.g., 1-10 uM) with varying concentrations of thiol-containing compounds (e.g., glutathione) in a buffer solution (e.g., PBS, pH 7.4) at room temperature. The fluorescence intensity (Ex/Em: 488/540 nm) is then measured with a spectrophotometer.
|
| Cell Assay |
A general protocol involves seeding cells in a confocal dish and incubating with Naph-EA-mal (e.g., 1-10 uM) for 10-30 minutes at 37degC. After washing, cells are fixed for protein thiol labeling or used live for thiol detection. Fluorescence imaging is performed with a confocal microscope (Ex/Em: 488/540 nm).
|
| Animal Protocol |
A typical protocol for in vivo imaging involves intravenous or intraperitoneal injection of an optimized concentration of Naph-EA-mal into a live animal model. After a suitable circulation time (e.g., 30 minutes to 2 hours), the animal is placed under an in vivo imaging system to monitor thiol-induced fluorescence.
|
| ADME/Pharmacokinetics |
General PK properties for small molecule probes are not applicable, as these compounds are typically used for acute imaging. However, formulation can be in PBS or saline with co-solvents like DMSO for in vivo use.
|
| Toxicity/Toxicokinetics |
General toxicity for imaging probes is considered low. For Naph-EA-mal, no specific toxicity studies are reported. In similar experiments, incubation with up to 10 uM for 24 hours in cell models shows good biocompatibility and no significant cytotoxicity.
|
| References | |
| Additional Infomation |
Naph-EA-mal is a research tool for studying redox biology, protein thiol modifications, and cellular redox states. Its ultrafast turn-on response and high specificity make it a powerful tool for real-time thiol detection. This product is for research use only.
|
| Molecular Formula |
C22H21N3O4
|
|---|---|
| Molecular Weight |
391.42
|
| Exact Mass |
391.153
|
| CAS # |
210292-65-2
|
| PubChem CID |
132564217
|
| Appearance |
Light yellow to orange solid powder
|
| LogP |
2.6
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
29
|
| Complexity |
716
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
N(C1=CC=C2C(N(CCCC)C(=O)C3C=CC=C1C2=3)=O)CCN1C(C=CC1=O)=O
|
| InChi Key |
RWRDYDLQZILWMW-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C22H21N3O4/c1-2-3-12-25-21(28)15-6-4-5-14-17(8-7-16(20(14)15)22(25)29)23-11-13-24-18(26)9-10-19(24)27/h4-10,23H,2-3,11-13H2,1H3
|
| Chemical Name |
2-butyl-6-[2-(2,5-dioxopyrrol-1-yl)ethylamino]benzo[de]isoquinoline-1,3-dione
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
DMSO: 33.33 mg/mL (85.15 mM)
|
|---|---|
| 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 | 2.5548 mL | 12.7740 mL | 25.5480 mL | |
| 5 mM | 0.5110 mL | 2.5548 mL | 5.1096 mL | |
| 10 mM | 0.2555 mL | 1.2774 mL | 2.5548 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
Copyright 2020 InvivoChem LLC | All Rights Reserved
