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Amplite® Fluorimetric Formaldehyde Quantitation Kit *Green Fluorescence*

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H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22


Formaldehyde is a naturally occurring substance. Natural processes in the upper atmosphere may contribute up to 90 percent of the total formaldehyde in the environment. Formaldehyde, as well as its oligomers and hydrates are rarely encountered in living organisms. Methanogenesis proceeds via the equivalent of formaldehyde, but this one-carbon species is masked as a methylene group in methanopterin. Formaldehyde is the primary cause of methanol's toxicity, since methanol is metabolized into toxic formaldehyde by alcohol dehydrogenase. Our Amplite® Fluorimetric Formaldehyde Quantitation Kit are used for quantifying formaldehyde. The kit uses a proprietary fluorogenic dye that generates a strongly fluorescent product upon reacting with formaldehyde. This fluorimetric kit provides a sensitive mix-and-read method to detect formaldehyde. The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation without a separation step. Its signal can be easily read with a fluorescence microplate reader.


Fluorescence microplate reader

Excitation410 nm
Emission525 nm
Cutoff495 nm
Recommended plateSolid black


Example protocol


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.


Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

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.


Formaldehyde standard

For convenience, use the Serial Dilution Planner: https://www.aatbio.com/tools/serial-dilution/10057

Add 12.2 µL of 123 mM Formaldehyde standard solution into 0.5 mL of Assay Buffer (Component B) to make 3 mM Formaldehyde standard solution. Take 3 mM Formaldehyde standard solution and perform 1:10 in Assay Buffer (Component B) to make 300 µM Formaldehyde standard (FS7).  Take 300 µM Formaldehyde standard  (FS7) and perform 1:3 serial dilutions to get serially diluted Formaldehyde standards (FS6-FS1) with Assay Buffer (Component B). 


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.

FS1 - FS750 µLSerial Dilutions (0.41 to 300 µM)
BL50 µLAssay Buffer
TS50 µLtest sample
  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).



View all 5 citations: Citation Explorer
High-Throughput Assay of Cytochrome P450-Dependent Drug Demethylation Reactions and Its Use to Re-evaluate the Pathways of Ketamine Metabolism
Authors: Davydova, Nadezhda Y and Hutner, David A and Gaither, Kari A and Singh, Dilip Kumar and Prasad, Bhagwat and Davydov, Dmitri R
Journal: (2023)
Biological Activity of Peptide-conjugated Polyion Complex Matrices Consisting of Alginate and Chitosan
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Journal: Peptide Science (2016)
Integrated self-assembling drug delivery system possessing dual responsive and active targeting for orthotopic ovarian cancer theranostics
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Journal: Biomaterials (2016): 12--26
Hepatic Deficiency of Augmenter of Liver Regeneration Exacerbates Alcohol-Induced Liver Injury and Promotes Fibrosis in Mice
Authors: Kumar, Sudhir and Wang, Jiang and Rani, Richa and G, undefined and hi, Ch and rashekhar R, undefined
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Fiber-optic protease sensor based on the degradation of thin gelatin films
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