Amplite® Fluorimetric Formaldehyde Quantitation Kit *Green Fluorescence*

Protocol summary

1. Prepare Formaldehyde standards and/or test samples (50 µL)
2. Add AldeLight™ Green working solution (50 µL)
3. Incubate at RT for 20 to 60 minutes
4. Monitor fluorescence increase at Ex/Em = 410/525 nm (Cutoff = 495 nm)

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

1. AldeLight™ Green stock solution (500X):
Add 20 µL of DMSO (Component D) into the vial of AldeLight™ Green (Component A) to make 500X AldeLight™ Green stock solution.

2. Formaldehyde stadard solution (123 mM):
Add 5 µL of 37.2% of Formaldehyde Standard (Component C) into 0.5 mL of Assay Buffer (Component B) to make 123 mM Formaldehyde standard solution.

Add 10 µL of 500X AldeLight™ Green stock solution into 5 mL of Assay Buffer (Component B) and mix well to make AldeLight™ Green working solution.  Note: 5 mL of AldeLight™ Green working solution is enough for 1 plate. AldeLight™ Green working solution is not stable, and best used within 2 hours.

Table 1. Layout of Formaldehyde standards and test samples in a solid back 96-well microplate. FS= Formaldehyde Standards (FS1 - FS7, 0.41 to 300 µM), BL=Blank Control, TS=Test Samples. 

Table 2. Reagent composition for each well.

1. Prepare Formaldehyde standards (FS), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2. For a 384-well plate, use 25 µL of reagent per well instead of 50 µL.
   
   
2. Add 50 µL of AldeLight™ Green working solution to each well of Formaldehyde standard, blank control, and test samples to make the total Formaldehyde assay volume of 100 µL/well. For a 384-well plate, add 25 µL of AldeLight™ Green working solution into each well instead, for a total volume of 50 µL/well.
   
   
3. Incubate the reaction at room temperature for 20 to 60 minutes, protected from light.
   
   
4. Monitor the fluorescence increase with a fluorescence plate reader at Ex/Em = 410/525 nm (Cutoff = 495nm).