Actively helping customers, employees and the global community during the coronavirus SARS-CoV-2 outbreak.  Learn more >>
Actively helping customers, employees and the global community during the coronavirus SARS-CoV-2 outbreak.  Learn more >>
logo
AAT Bioquest

Amplite® Fluorimetric Formaldehyde Quantitation Kit *Green Fluorescence*

Product Image
Product Image
Gallery Image 1
Ordering information
Price
Catalog Number
Unit Size
Quantity
Add to cart
Additional ordering information
Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12171501

OverviewpdfSDSpdfProtocol


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.

Platform


Fluorescence microplate reader

Excitation410 nm
Emission525 nm
Cutoff495 nm
Recommended plateSolid black

Components


Example protocol


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.

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: 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). 

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. 

BLBLTSTS
FS1FS1......
FS2FS2......
FS3FS3  
FS4FS4  
FS5FS5  
FS6FS6  
FS7FS7  

Table 2. Reagent composition for each well.

WellVolumeReagent
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).

Product Family


Images


Citations


View all 4 citations: Citation Explorer
Biological Activity of Peptide-conjugated Polyion Complex Matrices Consisting of Alginate and Chitosan
Authors: Fujimori, Chikara and Kumai, Jun and Nakamura, Kyotaro and Gu, Yingzi and Katagiri, Fumihiko and Hozumi, Kentaro and Kikkawa, Yamato and Nomizu, Motoyoshi
Journal: Peptide Science (2016)
Integrated self-assembling drug delivery system possessing dual responsive and active targeting for orthotopic ovarian cancer theranostics
Authors: Lin, Chun-Jui and Kuan, Chen-Hsiang and Wang, Li-Wen and Wu, Hsi-Chin and Chen, Yunching and Chang, Chien-Wen and Huang, Rih-Yang and Wang, Tzu-Wei
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
Journal: PloS one (2016): e0147864
Fiber-optic protease sensor based on the degradation of thin gelatin films
Authors: Schyrr, Bastien and Boder-Pasche, Stéphanie and Ischer, Réal and Smajda, Rita and Voirin, Guy
Journal: Sensing and Bio-Sensing Research (2015): 65--73

References


View all 23 references: Citation Explorer
Enzymatic method for formaldehyde assay based on formaldehyde dehydrogenase from the recombinant yeast Hansenula polymorpha
Authors: Demkiv OM, Haida HZ, Honchar MV.
Journal: Ukr Biokhim Zh (2009): 111
Intact and permeabilized cells of the yeast Hansenula polymorpha as bioselective elements for amperometric assay of formaldehyde
Authors: Khlupova M, Kuznetsov B, Demkiv O, Gonchar M, Csoregi E, Shleev S.
Journal: Talanta (2007): 934
The implausibility of systemic genotoxic effects measured by the comet assay in rats exposed to formaldehyde
Authors: Speit G., undefined
Journal: J Proteome Res (2006): 2523
Formaldehyde assay by capacitance versus voltage and impedance measurements using bi-layer bio-recognition membrane
Authors: Ben Ali M, Korpan Y, Gonchar M, El'skaya A, Maaref MA, Jaffrezic-Renault N, Martelet C.
Journal: Biosens Bioelectron (2006): 575
A new enzymo-chemical method for simultaneous assay of methanol and formaldehyde
Authors: Gonchar MV, Grabek D, Oklejewich B, Pavlishko HM, Shamlian OV, Sybirny VA, Kotylak Z, Rudke K, Csoregi E, Sibirny AA.
Journal: Ukr Biokhim Zh (2005): 146
Study on the formaldehyde-induced DNA damage with comet assay
Authors: Li R, Lu ZS, Qiao Y, Yao HC, Yu FF, Yang X.
Journal: Shi Yan Sheng Wu Xue Bao (2004): 262
Validation of an HPLC method on short columns to assay ketoconazole and formaldehyde in shampoo
Authors: Nguyen MN, Tallieu L, Plaizier-Vercammen J, Massart DL, V and er Heyden Y., undefined
Journal: J Pharm Biomed Anal (2003): 1
Quantification of bacterial lipopolysaccharides by the purpald assay: measuring formaldehyde generated from 2-keto-3-deoxyoctonate and heptose at the inner core by periodate oxidation
Authors: Lee CH, Tsai CM.
Journal: Anal Biochem (1999): 161
Assay of the inhibitory effect of pentachlorophenol and formaldehyde on mycelial growth and development of reproductive structures in Aspergillus niger
Authors: Bomar MT, Bomar M.
Journal: Folia Microbiol (Praha) (1999): 519
An enzyme-based formaldehyde assay and its utility in a sponge sterol biosynthetic pathway
Authors: Kerr RG, Kelly K.
Journal: J Nat Prod (1999): 201