Preparation of a copper-based fluorescent probe for nitric oxide and its use in mammalian cultured cells
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- Preparation of a copper-based fluorescent probe for nitric oxide and its use in mammalian cultured cells
- Lim, Mi Hee
- NOBEL LECTURE
- Issue Date
- NATURE PUBLISHING GROUP
- NATURE PROTOCOLS, v.2, no.2, pp.408 - 415
- A procedure for the preparation of a copper(II) complex (CuFL) as a fluorescent nitric oxide ( NO) detector is described. The fluorescein-based ligand FL can be synthesized in seven reaction steps (overall yield similar to 20%), typically requiring a total time of 9 days. The CuFL probe allows for the detection of NO produced in mammalian cultured cells. The detailed protocol for the use of CuFL for imaging NO in human neuroblastoma SK-N-SH cells takes a total time of similar to 26 h. This includes plating cells on six-well tissue culture plates or imaging dishes, treatment with CuFL, stimulation of NO synthases and imaging by fluorescence microscopy.
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