Amplite™ Fluorimetric Formaldehyde Quantitation Kit *Green Fluorescence*

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200 Tests 10057 $295

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Telephone: 1-800-990-8053
Fax: 1-408-733-1304
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Ex/Em (nm)400/510
Storage Freeze (<-15 °C)
Minimize light exposure
InstrumentsFluorescence microplate reader
Category Small Molecule Detection
Diagnostic Molecules
Related Redox Enzymes
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 at Ex/Em = 400/510 nm.


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This protocol only provides a guideline, and should be modified according to your specific needs.
At a glance

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.

Key parameters
Instrument:Fluorescence microplate reader
Excitation:410 nm
Emission:525 nm
Cutoff:495 nm
Recommended plate:Solid black
Preparation of stock solution
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.

Preparation of standard solution
Formaldehyde standard

For convenience, use the Serial Dilution Planner:

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). 

Preparation of working 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.

Sample experimental protocol

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. 

FS1 FS1 ... ...
FS2 FS2 ... ...
FS3 FS3    
FS4 FS4    
FS5 FS5    
FS6 FS6    
FS7 FS7    

Table 2. Reagent composition for each well.

Well Volume Reagent
FS1 - FS7 50 µL Serial Dilutions (0.41 to 300 µM)
BL 50 µL Assay Buffer
TS 50 µL test 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).
Example data analysis and figures

The reading (Response 1) obtained from the blank standard well is used as a negative control. Subtract this value from the other standards' readings to obtain the base-line corrected values. Then, plot the standards' readings to obtain a standard curve and equation. This equation can be used to calculate Concentration samples. We recommend using the Online Linear Regression Calculator which can be found at:

Figure 1.

AAT Bioquest provides high-quality reagents and materials for research use only. For proper handling of potentially hazardous chemicals, please consult the Safety Data Sheet (SDS) provided for the product. Chemical analysis and/or reverse engineering of any kit or its components is strictly prohibited without written permission from AAT Bioquest. Please call 408-733-1055 or email if you have any questions.

References & Citations

Biological Activity of Peptide-conjugated Polyion Complex Matrices Consisting of Alginate and Chitosan
Authors: Chikara Fujimori, Jun Kumai, Kyotaro Nakamura, Yingzi Gu, Fumihiko Katagiri, Kentaro Hozumi, Yamato Kikkawa, Motoyoshi Nomizu
Journal: Peptide Science (2016)

Hepatic Deficiency of Augmenter of Liver Regeneration Exacerbates Alcohol-Induced Liver Injury and Promotes Fibrosis in Mice
Authors: Sudhir Kumar, Jiang Wang, Richa Rani, Chandrashekhar R Gandhi
Journal: PloS one (2016): e0147864

Integrated self-assembling drug delivery system possessing dual responsive and active targeting for orthotopic ovarian cancer theranostics
Authors: Chun-Jui Lin, Chen-Hsiang Kuan, Li-Wen Wang, Hsi-Chin Wu, Yunching Chen, Chien-Wen Chang, Rih-Yang Huang, Tzu-Wei Wang
Journal: Biomaterials (2016): 12--26

Fiber-optic protease sensor based on the degradation of thin gelatin films
Authors: Bastien Schyrr, Stéphanie Boder-Pasche, Réal Ischer, Rita Smajda, Guy Voirin
Journal: Sensing and Bio-Sensing Research (2015): 65--73

Additional Documents

Safety Data Sheet (SDS)

Application Notes
1. AssayWise Letters 2013, Vol 2(1)
2. AssayWise Letters 2012, Vol 1(2)

Certificate of Analysis