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| Product Name | Fluorescein Diacetate (FDA) |
| Catalog No. | RESK-0331 |
| Description | Fluorescein diacetate (FDA) is a cell membrane-permeable esterase substrate commonly used for cell viability analysis. It can be used to measure esterase activity, because FDA must be catalytically activated by esterases to produce green fluorescence, and it can also be used to assess cell membrane integrity, as fluorescence is maintained only within intact membranes. It is also suitable for flow cytometric analysis. The maximum excitation (Ex) and emission (Em) wavelengths are 490 nm and 526 nm, respectively. Additional applications include: 1) studies of intracellular interactions; 2) use as a substrate for histochemical esterase detection; 3) studies of mitochondrial potential in isolated hepatocytes. Live cells with active esterases fluoresce green, whereas dead cells are not stained. |
| Intended Use | Cell viability staining and esterase activity measurement; assessment of cell membrane integrity; applicable to mammalian cells, bacteria, and yeast. |
| Principle / Technology | Non-fluorescent, membrane-permeant FDA enters cells and is hydrolyzed by intracellular esterases to fluorescein, which emits green fluorescence and is retained only in cells with intact membranes; live cells fluoresce green while dead cells remain unstained. |
| Detection Method | Fluorescence microscopy or flow cytometry |
| Sample Type | Mammalian cells, bacteria, yeast, and isolated hepatocytes. |
| Performance Range / Specifications | English synonyms: 3,6-Diacetoxyfluoran; Di-O-acetylfluorescein CAS NO.: 596-09-8 Molecular formula: C24H16O7 Molecular weight: 416.39 Melting point: 204-210 °C Purity: >98% (HPLC) Solubility: Soluble in chloroform, DMSO, and ethanol; slightly soluble in water Ex/Em: 490/526 nm |
| Specificity | Fluorescence requires intracellular esterase activity and an intact cell membrane; live cells are stained green and dead cells are not stained. |
| Reaction Conditions / Protocol | Example procedure for bacterial samples: 1. Prepare a 10 mM FDA stock solution: completely dissolve 4.16 mg of FDA in 1 mL of DMSO. 2. Dilute the FDA stock solution 1:800 into bacterial culture medium (fresh or overnight culture medium of the test bacteria) or PBS. 3. Dilute the bacteria with PBS to 10^5-10^7 cells/mL; filter 1 mL of the bacterial dilution through a 0.45 um filter to retain the bacteria. 4. Add 1 mL of FDA working solution and incubate at room temperature for 5-10 min before detection. |
| Components / Formulation | Fluorescein diacetate, supplied as solid powder. |
| Storage Conditions | Store at -20 °C, protected from light and kept dry. |
| Shelf Life | 2 years |
| Package Specifications | 1 g 5 g |
| Product Form | Solid powder |
| Purity | >98% (HPLC) |
| Concentration | 10 mM stock solution: 4.16 mg FDA dissolved in 1 mL DMSO. |
| Molecular Weight | 416.39 |
| Shipping Conditions | Shipped with ice packs. |
| Regulatory / Compliance | For research use only. Not for use in diagnostic or therapeutic procedures. |
| Recommended Buffer System | PBS or bacterial culture medium for working solution dilution; DMSO for stock solution preparation. |
| Application Notes / Precautions | Fluorescence is maintained only within cells with intact membranes. Protect from light during storage and handling. Reference for yeast/bacterial viability testing: PMID: 15362884. |
For research use only, not for clinical use.
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