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.
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.
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.
|Solubility (25°C)||DMSO ≥ 90 mg/mL|
|Storage||-20°C, protect from light|
|Body Surface Area (m2)||0.007||0.025||0.15||0.05||0.02||0.5|
|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.
 Kenji Yamatoya, et al. Regen Ther. Cryopreservation of undifferentiated and differentiated human neuronal cells
 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
 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
 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
 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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