| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| 100mg | |||
| Other Sizes |
| Targets |
CMFDA targets thiols (cysteine residues) in cells, forming a fluorescent adduct. It is cleaved by non-specific esterases common in living cells, producing a fluorescent compound, fluorescein. The compound also inhibits glutathione S-transferase omega 1 (GSTO1) with an IC50 of 51 nM. Its primary use is as a cell tracker dye for long-term monitoring of cell proliferation, migration, and survival.
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|---|---|
| ln Vitro |
CMFDA is a cell-permeable fluorescent probe that does not affect cell viability or proliferation. It is transformed intracellularly into a cell-impermeant, fluorescent product. The compound is brightly fluorescent with standard filters. As a GSTO1 inhibitor, it has an IC50 of 51 nM. These properties make it valuable for cell tracking studies, apoptosis assays, and cellular assays.
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| ln Vivo |
In vivo, CMFDA is used for cell tracking and labeling in animal models. Cells labeled with CMFDA can be tracked after injection into animals to study cell migration, homing, and engraftment. The compound's fluorescence allows for visualization of labeled cells in tissues.
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| Enzyme Assay |
In vitro assays for CMFDA involve measuring its fluorescence properties and its inhibition of GSTO1. For GSTO1 inhibition, the enzyme is incubated with a substrate (e.g., glutathione and a thiol-reactive compound) in the presence of varying concentrations of CMFDA. Enzyme activity is measured spectrophotometrically, and IC50 values are calculated.
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| Cell Assay |
Cellular assays for CMFDA are performed by incubating cells with the dye at concentrations of 1-10 μM for 15-60 minutes. After washing, cells are analyzed by flow cytometry or fluorescence microscopy. The dye is retained in cells for several days, allowing for long-term tracking. Cell viability is assessed to confirm that the dye does not affect cell health.
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| Animal Protocol |
In vivo animal studies with CMFDA involve labeling cells ex vivo and then injecting them into animals. Labeled cells are tracked by fluorescence imaging or by harvesting tissues and analyzing them by flow cytometry or fluorescence microscopy. The dye's retention in cells allows for long-term tracking of cell migration and engraftment.
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| ADME/Pharmacokinetics |
CMFDA is cell-permeable and retained in cells after intracellular conversion. Its molecular weight is 464.86 g/mol. The compound is typically dissolved in DMSO for stock solutions and diluted in cell culture medium for use. It is stable in cells for several days.
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| Toxicity/Toxicokinetics |
CMFDA is generally non-toxic and does not affect cell viability or proliferation. No significant toxicity has been reported at the concentrations used for cell labeling (1-10 μM). The compound is intended for research use and should be handled with standard safety precautions.
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| References | |
| Additional Infomation |
5-Chloromethylfluorescein diacetate is an acetate. It is a fluorescent dye. Functionally, it is related to fluorescein (the lactone form).
CMFDA is a widely used fluorescent cell tracker dye for studying cell proliferation, migration, and survival. It is also a potent GSTO1 inhibitor. The compound is available from multiple suppliers for research use. It is not approved for clinical use and is intended for laboratory research only. |
| Molecular Formula |
C25H17O7CL
|
|---|---|
| Molecular Weight |
464.85128
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| Exact Mass |
464.066
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| CAS # |
136832-63-8
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| PubChem CID |
122297
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| Appearance |
White to off-white solid powder
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| Density |
1.49 g/cm3
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| Boiling Point |
657.5ºC at 760 mmHg
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| Flash Point |
238.3ºC
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| Vapour Pressure |
3.65E-17mmHg at 25°C
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| Index of Refraction |
1.675
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| LogP |
4
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
33
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| Complexity |
760
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC(=O)OC1=CC2=C(C=C1)C3(C4=C(C=C(C=C4)CCl)C(=O)O3)C5=C(C=C(C=C5)OC(=O)C)O2
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| InChi Key |
IPJDHSYCSQAODE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C25H17ClO7/c1-13(27)30-16-4-7-20-22(10-16)32-23-11-17(31-14(2)28)5-8-21(23)25(20)19-6-3-15(12-26)9-18(19)24(29)33-25/h3-11H,12H2,1-2H3
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| Chemical Name |
[6'-acetyloxy-5-(chloromethyl)-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl] acetate
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| Synonyms |
CM-FDA; Cell Tracker Green; CMFDA
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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 : ~33.33 mg/mL (~71.70 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 1.43 mg/mL (3.08 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 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 14.3 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. Solubility in Formulation 2: ≥ 1.43 mg/mL (3.08 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 14.3 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1512 mL | 10.7562 mL | 21.5123 mL | |
| 5 mM | 0.4302 mL | 2.1512 mL | 4.3025 mL | |
| 10 mM | 0.2151 mL | 1.0756 mL | 2.1512 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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