4-amino-5-methylamino-2 ,7 -difluorofluorescein

Sheng, J. Wang, D. Braun, A. P. DAF-FM (4-amino-5-methylamino-2, 7 -difluorofluorescein) diacetate detects impairment of agonist-stimulated nitric oxide synthesis by elevated glucose in human vascular endothelial cells Reversal by vitamin C and L-sepiapterin. J. Pharmacol. Exp. Then 2005, 315, 931-940. 

Nitric oxide is a by-product of the action of the enzyme NO-synthetase on L-aiginine in tissues. The NO product is reacted with 3-amino-4-(N-methylamino)-2, 7 -difluorofluorescein and analyzed on a 30 C C,g column (A=500nm, ex 515nm, em) using a 94/6 water (lOmM Na2HP04 buffer at pH 7.2)/acetonitrile mobile phase [906]. Elution was complete in 14 min. A linear range of 2-200 nM was claimed. 

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