5-CFDA

• CAT Number: R055467
• CAS Number: 79955-27-4
• Molecular Formula: C25H16O9
• Molecular Weight: 460.39
• Purity: ≥95%
• Synonyms: 3′,6′-diacetyloxy-3-oxospiro[2-benzofuran-1,9′-xanthene]-5-carboxylic acid
• InChI: InChI=1S/C25H16O9/c1-12(26)31-15-4-7-19-21(10-15)33-22-11-16(32-13(2)27)5-8-20(22)25(19)18-6-3-14(23(28)29)9-17(18)24(30)34-25/h3-11H,1-2H3,(H,28,29)
• InChIKey: WPUZGNPQMIWOHE-UHFFFAOYSA-N
• SMILES: CC(=O)OC1=CC2=C(C=C1)C3(C4=C(C=C(C=C4)C(=O)O)C(=O)O3)C5=C(O2)C=C(C=C5)OC(=O)C

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. CF containing portion contains an additional negative charge so that it is better retained in cells, compared to fluorescein dyes[1][2][3].
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. This product is for R&D use only, not for drug, household, or other uses. 4. For your safety and health, please wear a lab coat and disposable gloves to operate.

Reference

[1]. Yang T, et al. A novel nonradioactive CFDA assay to monitor the cellular immune response in myeloid leukemia. Immunobiology. 2013 Apr;218(4):548-53.
 [Content Brief]

[2]. Card SD, et al. Assessment of fluorescein-based fluorescent dyes for tracing Neotyphodium endophytes in planta. Mycologia. 2013 Jan-Feb;105(1):221-9.
 [Content Brief]

[3]. Fang X, et al. Bone marrow-derived endothelial progenitor cells are involved in aneurysm repair in rabbits. J Clin Neurosci. 2012 Sep;19(9):1283-6.
 [Content Brief]