Amplite® Fluorimetric Fluorescamine Protein Quantitation Kit *Blue Fluorescence*

Protocol summary

1. Prepare  fluorescamine working solution (25 µL)
2. Add BSA standards or test samples (75 µL)
3. Incubate at room temperature for 5 - 30 minutes
4. Read fluorescence intensity at Ex/Em = 380/470 nm

Important notes
Thaw all the kit components at room temperature before starting the experiment.

Add the whole content of DMSO (Component B) into the bottle of Fluorescamine (Component A), and mix well. Note: 2.5 mL of fluorescamine working solution is enough for 1 plate.

Table 1. Layout of BSA standards and test samples in a solid black 96-well microplate. BS= BSA Standards (BS1 - BS7, 1.563 to 100 µg/mL), BL=Blank Control, TS=Test Samples.

Table 2. Reagent composition for each well.

1. Prepare BSA standards (BS), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2. For a 384-well plate, use 30 µL of reagent per well instead of 75 µL.
   
   
2. Add 25 µL of  fluorescamine working solution to each well of BSA standard, blank control, and test samples to make the total assay volume of 100 µL/well. For a 384-well plate, add 10 µL of fluorescamine working solution into each well instead, for a total volume of 40 µL/well.
   
   
3. Incubate the reaction at room temperature for 5 to 30 minutes, protected from light.
   
   
4. Monitor the fluorescence increase with a fluorescence plate reader at Ex/Em = 380/470 nm.