2,3-Diaminonaphthalene

2,3-Diaminonaphthalene is a fluorescent reagent used to detect selenium and nitric oxide (NO). 2,3-Diaminonaphthalene binds to selenium to form pyrazolene selenol, with excitation/emission maxima at 365 nm and 525 nm, respectively. 2,3-Diaminonaphthalene binds to NO2− to form the fluorescent product, 1-(H)-napthotriazole, with excitation/emission maxima at 360 nm and 440 nm, respectively^[1][2]. In Vitro:Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
Nitric oxide (NO) Assay^[2]
1. 2,3-Diaminonaphthalene was dissolved in 0.62 N HCl at a concentration of 0.05 mg/mL.
2. Aliquots of media (100 μL) were placed into 96-well plates in triplicate.
Note: Media were void of any interfering components such as dithiothreitol, protein, FBS, phenol red, or hemoglobin.
3. 2,3-Diaminonaphthalene (10 μL) was added to each well for 10 min at room temperature.
4. After 10 min, 5 μL 2.8 N NaOH were added to each well.
5. The plate was read on a luminescence spectrometer (Ex/Em = 360 nm/440 nm).