Fluorescamine is not fluorescent itself, but reacts with primary amines at alkaline pH to yield highly fluorescent products with Ex=365 nm and Em=470 nm.

Store at -20°C in dark

Protocol for Fluorescamin -based protein assay 1. Prepare a fresh solution of Fluorescamine at concentration of 1.08 mM in acetone. Keep in dark. 2. Prepare 2-fold serial dilutions of BSA standard (final concentration 0-1000 μg, or as indicated) dissolved in PBS to a final volume of 200 μL in 96-well plate. This is prepared for a standard curve. 3. Prepare the sample dissolved in PBS to a final volume of 200 μL at the same plate. 4. Add 50 μL Fluorescamin solution to every well. Shake the plate to mix the solution well. 5. Incubate for 15 min at room temperature in dark. 6. Read the plate on multi-well fluorescence plate reader (400 nm excitation filter, half bandwidth 35 nm; 460 nm emission filter, half bandwidth 40 nm). 7. To obtain the sample protein concentrations, the fluorescent intensity data are calibrated against the BSA standard curve.{{Lorenzen A, Kennedy SW. A fluorescence-based protein assay for use with a microplate reader. Anal Biochem. 1993 Oct;214(1):346-8. doi: 10.100

Fluorescamine is not fluorescent itself, but reacts with primary amines at alkaline pH to yield highly fluorescent products with Ex=365 nm and Em=470 nm.