| Size | Price | Stock | Qty |
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| 5mg |
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| Targets |
Fluorescent Dye; Emission (Em) = 515; Excitation (Ex) = 419
Calcein tetraethyl ester does not have a specific biological target as a therapeutic agent. It is a fluorescent probe designed for cell viability and cytotoxicity assays. The tetraethyl ester groups enable cell permeability by masking the negative charges of the carboxyl groups. Once inside the cell, esterases cleave the ester groups, converting the compound into the charged, membrane-impermeant fluorescent dye calcein, which remains trapped inside viable cells. Calcein exhibits green fluorescence, allowing detection of live cells. |
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| ln Vitro |
Calcein tetraethyl ester is a cell-permeable fluorescent dye widely used in cell viability and cytotoxicity assays. It can enter living cells through the cell membrane. Once inside live cells, it is converted by intracellular esterases into the fluorescent dye calcein, which is trapped inside viable cells and exhibits green fluorescence. The compound is particularly useful in live/dead assays (often used in combination with propidium iodide or ethidium homodimer), calcium flux studies, and for monitoring intracellular processes. It provides a sensitive and non-toxic method for evaluating the effects of experimental treatments.
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| ln Vivo |
No in vivo activity data are documented for Calcein tetraethyl ester as a therapeutic agent. The compound is a research tool used for cell viability and cytotoxicity assays in isolated cells. It is not intended for therapeutic applications. In vivo applications are limited due to rapid hydrolysis and clearance. The compound is primarily used in ex vivo and in vitro experimental settings.
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| Enzyme Assay |
In vitro enzyme assays for Calcein tetraethyl ester are not typically performed as it is not an enzyme inhibitor. The compound is used as a substrate for esterases, which cleave the ester groups to release the fluorescent calcein. For quality control and characterization, the hydrolysis of Calcein tetraethyl ester by esterases can be monitored by fluorescence spectroscopy. The increase in fluorescence at 530 nm upon excitation at 490 nm indicates successful de-esterification. The compound is stable in dry form and should be protected from moisture and light.
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| Cell Assay |
Cell-based assays with Calcein tetraethyl ester are performed to assess cell viability and cytotoxicity. Cells are incubated with Calcein tetraethyl ester (typically 1-10 μM) in culture medium for 15-60 minutes at 37°C. The ester groups are cleaved by intracellular esterases in live cells, trapping calcein inside the cells. Fluorescence is measured using a fluorescence microscope, plate reader, or flow cytometer (excitation 490 nm, emission 530 nm). Live cells show green fluorescence, while dead cells do not (as they lack esterase activity or cannot retain the dye). The compound can be used in combination with a dead-cell stain (e.g., propidium iodide or ethidium homodimer-1) for live/dead discrimination.
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| Animal Protocol |
No animal models are documented for Calcein tetraethyl ester as it is a fluorescent dye rather than a therapeutic agent. The compound is used for ex vivo measurements in isolated tissues or cells. For certain research applications, the dye may be injected into animals for in vivo imaging, but its rapid hydrolysis and clearance limit its utility. The compound is primarily used in cell-based and tissue-based assays for viability and cytotoxicity assessment.
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| ADME/Pharmacokinetics |
Calcein tetraethyl ester has a molecular weight of 734.75 and molecular formula C38H42N2O13. It has a melting point of 146-151°C and a boiling point of 825.4°C (predicted). The compound is light-sensitive and should be protected from light during handling and storage. It is soluble in DMSO and should be stored as a stock solution in DMSO at -20°C, protected from light and moisture. The compound is hygroscopic and should be handled under dry conditions. Solutions should be prepared fresh before use.
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| Toxicity/Toxicokinetics |
No specific toxicity data are documented for Calcein tetraethyl ester. The compound is a fluorescent dye used in cell biology research and is generally considered non-toxic at the concentrations used for viability assays (1-10 μM). It provides a non-toxic method for monitoring intracellular processes. Standard laboratory safety practices should be followed when handling the compound. The compound is intended for research use only and is not approved for human therapeutic applications.
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| Additional Infomation |
Calcein tetraethyl ester is also known as fluorexon tetraethyl ester. It is a cell-permeable fluorescent dye widely used in cell viability and cytotoxicity assays. The compound can enter living cells and is converted by esterases into the fluorescent dye calcein, which is trapped inside viable cells. It is commonly used in live/dead assays (with propidium iodide or ethidium homodimer), calcium flux studies, and for monitoring intracellular processes. The compound provides a sensitive and non-toxic method for evaluating experimental treatments. It is also an intermediate in the synthesis of calcein AM. It is intended for research use only.
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| Molecular Formula |
C38H42N2O13
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|---|---|
| Molecular Weight |
734.745691776276
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| Exact Mass |
734.268
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| CAS # |
1170856-93-5
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| Related CAS # |
Calcein;1461-15-0
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| PubChem CID |
91810997
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| Appearance |
Light yellow to yellow solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
825.4±65.0 °C at 760 mmHg
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| Flash Point |
453.0±34.3 °C
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| Vapour Pressure |
0.0±3.1 mmHg at 25°C
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| Index of Refraction |
1.638
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| LogP |
6.56
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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 |
20
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| Heavy Atom Count |
53
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| Complexity |
1180
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCOC(=O)CN(CC1=C(C=CC2=C1OC3=C(C24C5=CC=CC=C5C(=O)O4)C=CC(=C3CN(CC(=O)OCC)CC(=O)OCC)O)O)CC(=O)OCC
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| InChi Key |
AWKMLDYACNCHOZ-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C38H42N2O13/c1-5-48-31(43)19-39(20-32(44)49-6-2)17-24-29(41)15-13-27-35(24)52-36-25(18-40(21-33(45)50-7-3)22-34(46)51-8-4)30(42)16-14-28(36)38(27)26-12-10-9-11-23(26)37(47)53-38/h9-16,41-42H,5-8,17-22H2,1-4H3
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| Chemical Name |
ethyl 2-[[5'-[[bis(2-ethoxy-2-oxoethyl)amino]methyl]-3',6'-dihydroxy-3-oxospiro[2-benzofuran-1,9'-xanthene]-4'-yl]methyl-(2-ethoxy-2-oxoethyl)amino]acetate
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| Synonyms |
Calcein tetraethyl ester; 1170856-93-5; Calcein (tetraethyl ester); Fluorexon tetraethyl ester; tetraethyl 2,2',2'',2'''-(((3',6'-dihydroxy-3-oxo-3H-spiro[isobenzofuran-1,9'-xanthene]-4',5'-diyl)bis(methylene))bis(azanetriyl))tetraacetate; ethyl 2-[[5'-[[bis(2-ethoxy-2-oxoethyl)amino]methyl]-3',6'-dihydroxy-3-oxospiro[2-benzofuran-1,9'-xanthene]-4'-yl]methyl-(2-ethoxy-2-oxoethyl)amino]acetate; Glycine, N,N'-[(3',6'-dihydroxy-3-oxospiro[isobenzofuran-1(3H),9'-[9H]xanthene]-4',5'-diyl)bis(methylene)]bis[N-(2-ethoxy-2-oxoethyl)-, 1,1'-diethyl ester; Tetraethyl Fluorescein Bis(methylene)bis(azanetriyl)tetraacetate;
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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: 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)
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| Solubility (In Vitro) |
DMSO : ~25 mg/mL (~34.03 mM)
H2O : ~0.67 mg/mL (~0.91 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (3.40 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3610 mL | 6.8050 mL | 13.6101 mL | |
| 5 mM | 0.2722 mL | 1.3610 mL | 2.7220 mL | |
| 10 mM | 0.1361 mL | 0.6805 mL | 1.3610 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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