Kinase experiment:

Various concentrations of each drug are incubated for 60 min at 37℃ in OS-mL reaction volumes containing PBS (pH 7.4), 200 μM 2,3-Diaminonaphthalene (DAN) and 40 μM Sp/NO. Spermine/NO will spontaneously decompose at pH7.4 to produce 2 mol of NO and 1 mol of spermine with a half-life of 39 min at 37℃. Following the incubation period, 2.5 mL of 10 mM NaOH is added to each tube to stop the reaction. The NO-dependent N-nitrosation of 2,3-Diaminonaphthalene to yield its highly fluorescent N-nitrosated derivative, 2,3-naphthotriazole, is quantified by measuring the fluorescence of each sample using an excitation wavelength of 375 nm and an emission wavelength of 450nm. The concentration of Triazole is determined using standards of the pure 2,3-naphthotriazole[3].

2,3 Diaminonaphthalene is a highly selective colorimetric and fluorometric reagent for selenium detection and also used for the fluorometric determination of nitrite.

Poly(2,3-diaminonaphthalene) microspheres as a novel quencher for fluorescence-enhanced nucleic acid detection[1]. 2,3-Diaminonaphthalene (DAN) is possibly a carcinogenic reagent[2].

[1]. Tian J, et al. Poly(2,3-diaminonaphthalene) microspheres as a novel quencher for fluorescence-enhanced nucleic acid detection. Analyst. 2011 Jun 7;136(11):2221-4. [2]. Martínez-Tomé MJ, et al. Immobilization and characterization of 2,3-diaminonaphthalene/cyclodextrin complexes in a sol-gel matrix: a new fluorimetric sensor for nitrite. J Fluoresc. 2009 Jan;19(1):119-25. [3]. Grisham MB, et al. Effects of aminosalicylates and immunosuppressive agents on nitric oxide-dependent N-nitrosation reactions. Biochem Pharmacol. 1994 May 18;47(10):1897-902.