Amplite® Colorimetric Aldehyde Quantitation Kit

Rapid and accurate measurement of aldehydes is important task for biological research, food industry, chemical research and environmental pollution surveillance. There are few reagents or assay kits available for quantifying the amount of aldehydes. Most of existing aldehyde test methods is based on separations either by the tedious and expensive HPLC-MS or GC-MS. Our Amplite® Colorimetric Aldehyde Quantitation kit uses a proprietary dye that generates a chromogenic product upon reacting with an aldehyde. The kit provides a sensitive, one-step colorimetric method to detect as little as 1 nanomole of aldehyde in a 100 µL assay volume (10 µM; Figure 1). The assay can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation with no separation steps required. Its signal can be easily read by absorbance microplate reader at 550 nm.

Example protocol

AT A GLANCE

Protocol Summary

Prepare Aldehyde standards and/or test samples (50 µL)
Add 2X AldeView™ Yellow working solution (50 µL)
Incubate at room temperature for 30 to 60 minutes
Monitor absorbance increase at 550 nm

Important Thaw all the kit components to room temperature before starting the experiment. Assay solution (Component B) is potentially hazardous. Wear gloves when handling it.

PREPARATION OF STOCK SOLUTIONS

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.

Aldehyde standard solution (10 mM)

Add 1 mL of Dilution Buffer (Component D) into the vial of Aldehyde Standard (Component C) to make a 10 mM aldehyde standard solution.

PREPARATION OF STANDARD SOLUTION

For convenience, use the Serial Dilution Planner:
https://www.aatbio.com/tools/serial-dilution/10051

Aldehyde standard

Take 10 mM Aldehyde standard solution and perform 1:10 dilution to get 1000 uM Aldehyde standard solution (AS7). Then perform 1:3 serial dilutions to get remaining serial dilutions of aldehyde standard (AS6 - AS1).

PREPARATION OF WORKING SOLUTION

Add 5 mL of Assay Solution (Component B) into the bottle of AldeView™ Yellow (Component A), and mix well.
Note 5 mL of the 2X AldeView™ Yellow reaction mixture is enough for 1 plate. The reaction mixture is not stable. Use within 2 hours.
Note Assay solution (Component B) is potentially hazardous. Wear gloves when handling it.

SAMPLE EXPERIMENTAL PROTOCOL

Table 1. Layout of Aldehyde standards and test samples in a white/clear bottom 96-well microplate. AS= Aldehyde Standards (AS1 - AS7, 1 to 1000 µM), BL=Blank Control, TS=Test Samples.

BL	BL	TS	TS
AS1	AS1	...	...
AS2	AS2	...	...
AS3	AS3
AS4	AS4
AS5	AS5
AS6	AS6
AS7	AS7

Table 2.Reagent composition for each well.

Well	Volume	Reagent
AS1 - AS7	50 µL	Serial Dilution (1 to 1000 µM)
BL	50 µL	Assay Buffer
TS	50 µL	test sample

Prepare aldehyde standards (AS), 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. Note: Both BSA and Tween 20 will interfere with the assay, use less than 0.001% BSA and 0.01% Tween 20 in the samples.
Note If the aldehyde-containing samples are from enzyme reactions such as fructose-1,6-bisphosphate with fructose-1,6-bisphosphate aldolase, prepare 50 µL of enzyme reaction (25 µL for a 384-well plate) as desired. Incubate the enzyme reaction at 37 °C for at least 1 hour. The components of enzyme reaction should be optimized as needed (e.g. an optimized buffer system might be required for a specific enzyme reaction). In most cases, Dilution Buffer (Component D) can also be used for running enzyme reaction if you do not have an optimized enzyme buffer.
Add 50 µL of AldeView™ Yellow working solution to each well of aldehyde standard, blank control, and test samples to make the total aldehyde assay volume of 100 µL/well. For a 384-well plate, add 25 µL of AldeView™ Yellow working solution into each well instead, for a total volume of 50 µL/well.
Incubate the reaction mixture at room temperature for 30 to 60 minutes, protected from light.
Monitor the absorbance increase with an absorbance plate reader at 550 nm.
Note Different concentrations of the aldehyde might form different colors with AldeView™ Yellow.

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 19 references: Citation Explorer

Measurement of atmospheric hydroxyacetone, glycolaldehyde, and formaldehyde
Authors: Zhou X, Huang G, Civerolo K, Schwab J.
Journal: Environ Sci Technol (2009): 2753

Formaldehyde cross-linking of proteins to osteoclast differentiation factor promoter for the identification of biofunctional proteins
Authors: Haghighi M., undefined
Journal: Saudi Med J (2009): 472

Methods to reduce formaldehyde levels of cadavers
Authors: Whitehead MC., undefined
Journal: Clin Anat (2009): 421

Formaldehyde in China: production, consumption, exposure levels, and health effects
Authors: Tang X, Bai Y, Duong A, Smith MT, Li L, Zhang L.
Journal: Environ Int (2009): 1210

Urine formaldehyde level is inversely correlated to mini mental state examination scores in senile dementia
Authors: Tong Z, Zhang J, Luo W, Wang W, Li F, Li H, Luo H, Lu J, Zhou J, Wan Y, He R.
Journal: Neurobiol Aging. (2009)

Page updated on November 21, 2024

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