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5-CFDA

Cat. No. M21716
5-CFDA Structure
Synonym:

5-Carboxyfluorescein diacetate

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Biological Activity

5-CFDA is a common aliphatic luciferin-line organism. CFDA conducts free diffusion into cells, and then it is hydrolyzed into carboxyl fluorescein (CF) by intracellular non-specific lipase.

Preparation of 5-CFDA working solution

1.Preparation of the stock solution
Dissolve 1mg 5-CFDA in 0.2172 mL DMSO to obtain 10 mM of 5-CFDA.
Note: It is recommended to store the stock solution at -20°C -80°C away from light and avoid repetitive freeze-thaw cycles.

2.Preparation of 5-CFDA working solution
Dilute the stock solution in serum-free cell culture medium or PBS to obtain 1-10 μM of 5-CFDA working solution.
Note: Please adjust the concentration of 5-CFDA working solution according to the actual situation.

Cell staining

1.Cell preparation:
For suspension cells: Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.
For adherent cells: Discard the cell culture medium, and add trypsin to dissociate cells to make a single-cell suspension. Centrifuge at 1000 g at 4°C for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.

2.Add 1 mL of 5-CFDA working solution, and then incubate at room temperature for 30 minutes.

3.Centrifuge at 400 g at 4°C for 3-4 minutes and then discard the supernatant.

4.Wash twice with PBS, 5 minutes each time.

5.Resuspend cells with serum-free cell culture medium or PBS, and then detect by fluorescence microscope or flow cytometer.

Precautions

1. It is recommended to store the stock solution at -20℃ or -80℃ away from light and avoid repetitive freeze-thaw cycles.

2. Please adjust the concentration of 5-CFDA working solution according to the actual situation.

3. For your safety and health, please wear a lab coat and disposable gloves to operate.

Chemical Information
Molecular Weight 460.39
Formula C25H16O9
CAS Number 79955-27-4
Solubility (25°C) DMSO ≥ 90 mg/mL
Storage -20°C, protect from light
Conversion of different model animals based on BSA (PMID: 27057123)
Species Mouse Rat Rabbit Guinea pig Hamster Dog
Weight (kg) 0.02 0.15 1.8 0.4 0.08 10
Body Surface Area (m2) 0.007 0.025 0.15 0.05 0.02 0.5
Km factor 3 6 12 8 5 20
Animal A (mg/kg) = Animal B (mg/kg) multiplied by  Animal B Km
Animal A Km

For example, to modify the dose of Compound A used for a mouse (20 mg/kg) to a dose based on the BSA for a rat, multiply 20 mg/kg by the Km factor for a mouse and then divide by the Km factor for a rat. This calculation results in a rat equivalent dose for Compound A of 10 mg/kg.

References

[1] Kenji Yamatoya, et al. Regen Ther. Cryopreservation of undifferentiated and differentiated human neuronal cells

[2] Fatemeh Mosaffa, et al. Iran J Basic Med Sci. Synthesis and biological evaluation of novel quinoline analogs of ketoprofen as multidrug resistance protein 2 (MRP2) inhibitors

[3] T Jambor, et al. Physiol Res. Investigation of the Properties and Effects of Salvia Officinalis L. on the Viability, Steroidogenesis and Reactive Oxygen Species (ROS) Production in TM3 Leydig Cells in Vitro

[4] Elena Gorokhova, et al. J Microbiol Methods. A comparison of TO-PRO-1 iodide and 5-CFDA-AM staining methods for assessing viability of planktonic algae with epifluorescence microscopy

[5] M E Fuller, et al. Appl Environ Microbiol. Development of a vital fluorescent staining method for monitoring bacterial transport in subsurface environments

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