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| Product Name | Green Cell Tracking Dye (CMFDA) |
| Catalog No. | RESK-0332 |
| Description | CMFDA (5-chloromethylfluorescein diacetate) is a fluorescent dye that freely permeates live cell membranes for cell tracking. CMFDA is a chloromethyl derivative of fluorescein diacetate (FDA); it is cell membrane-permeable and non-fluorescent. After entering live cells by passive transport, its lipophilic groups are hydrolyzed by nonspecific cytoplasmic esterases to generate 5-chloromethylfluorescein, which emits green fluorescence. Because it carries a charge, the product cannot freely cross the cell membrane, and its chloromethyl group forms adducts with glutathione in intracellular proteins and polypeptides under the action of glutathione S-transferase, so it is well retained inside the cell. Labeled cells show very stable fluorescence, with stable labeling lasting for days (at least 3 days), making the dye highly suitable for studies of cell differentiation and morphogenesis; compared with commonly used calcein and FDA, it offers longer intracellular signal retention. During cell division and proliferation, the CMFDA label is distributed to both daughter cells without staining neighboring cells (staining via gap junctions occurs only in rare cases). CMFDA labeling is also used to analyze cell-cell fusion, cell adhesion, and multidrug resistance transporters. In addition, CMFDA can be combined with other cell tracking dyes to selectively label pathogens within infected cells, and used together with membrane-impermeant nuclear fluorescent probes for cell viability studies. This product has the ideal properties of a cell tracking reagent: stable, non-toxic within the working concentration range, well retained intracellularly, and strongly fluorescent over the physiological pH range. CMFDA-labeled cells show green fluorescence (Ex = 492 nm, Em = 517 nm) and can be co-stained with fluorophores of other colors, such as red fluorescent protein (RFP). The product is supplied as a powder and must be prepared as a stock solution in DMSO before use. |
| Intended Use | Long-term fluorescent tracking of live cells, including cell differentiation, morphogenesis, proliferation, cell fusion, adhesion, and multidrug resistance transporter studies. |
| Principle / Technology | Membrane-permeant, non-fluorescent CMFDA is hydrolyzed by intracellular esterases to green-fluorescent 5-chloromethylfluorescein; the chloromethyl group covalently couples to intracellular thiols (glutathione) via glutathione S-transferase, forming membrane-impermeant adducts that are stably retained in the cell. |
| Detection Method | Fluorescence microscopy / fluorescence imaging |
| Sample Type | Live animal and plant cells (lipophilic dye with no cell-type specificity). |
| Performance Range / Specifications | English synonym: 5-Chloromethylfluorescein diacetate CAS NO.: 136832-63-8 Molecular formula: C25H17ClO7 Molecular weight: 464.85 Appearance: White or off-white solid Ex/Em: 492/517 nm |
| Specificity | No cell-type specificity; labels live animal and plant cells. |
| Reaction Conditions / Protocol | Prepare a stock solution in DMSO before use, then dilute to a working solution for cell staining. Recommended staining volume: 500 uL of staining working solution per 200,000-1,000,000 cells. |
| Components / Formulation | CMFDA (5-chloromethylfluorescein diacetate), supplied as a white or off-white solid powder. |
| Storage Conditions | Store at -20 °C, dry and protected from light. Avoid repeated freeze-thaw cycles. |
| Shelf Life | 1 year |
| Package Specifications | 1 x 50 ug 2 x 50 ug 5 x 50 ug |
| Product Form | White or off-white solid powder |
| Molecular Weight | 464.85 |
| pH Range / Optimal pH | Strong fluorescence over the physiological pH range. |
| Shipping Conditions | Shipped at room temperature. |
| Regulatory / Compliance | For research use only. Not for use in diagnostic or therapeutic procedures. |
| Compatibility | Can be co-stained with fluorophores of other colors, such as red fluorescent protein (RFP); can be combined with other cell tracking dyes to selectively label pathogens in infected cells, and used with membrane-impermeant nuclear probes for viability studies. |
| Recommended Buffer System | DMSO for stock solution preparation. |
| Application Notes / Precautions | Fluorescent probes have some toxicity to live cells; complete detection and observation as soon as possible after staining. Minimize the time between probe loading and measurement (excluding stimulation time). If the fluorescence signal is weak, increase the staining time and/or concentration. Avoid repeated freeze-thaw cycles. |
For research use only, not for clinical use.
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