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Amplite® Colorimetric Aldehyde Quantitation Kit *Blue Color*
Example protocol
AT A GLANCE
Protocol summary
- Prepare Aldehyde standards and/or test samples (50 µL)
- Add 2X AldeView™ Blue working solution (50 µL)
- Incubate at RT for 20 minutes
- Add AldeView™ Blue Enhancer (50 µL)
- Incubate at RT for 20 minutes
- Monitor absorbance increase at 620 nm
Important notes
Thaw all the kit components to room temperature before starting the experiment.
PREPARATION OF STOCK SOLUTION
1. Aldehyde standard solution (10 mM):
Add 1 mL of Assay Buffer (Component B) into the vial of Aldehyde Standard (Component D) 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/10053
Take 10 mM Aldehyde standard solution and perform 1:100 in Assay Buffer (Component B) to make 100 µM Aldehyde standard solution (AS7). Take 100 µM Aldehyde standard solution (AS7) and perform 1:2 serial dilutions to get serially diluted Aldehyde standards (AS6 - AS1) with Assay Buffer (Component B).
PREPARATION OF WORKING SOLUTION
Add 5 mL of Assay Buffer (Component B) into one bottle of AldeView™ Blue (Component A) to make 2X AldeView™ Blue working solution. Note: 5 mL of 2X AldeView™ Blue working solution is enough for one plate. 2X AldeView™ Blue working solution is not stable, and best used within 2 hours.
SAMPLE EXPERIMENTAL PROTOCOL
Table 1. Layout of Aldehyde Standards and test samples in a white 96-well microplate with clear bottom. AS= Aldehyde Standards (AS1 - AS7, 1.56 to 100 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 Dilutions (1.56 to 100 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 12.5 µL of reagent per well instead of 50 µL.
- Add 50 µL of AldeView™ Blue 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 12.5 µL of AldeView™ Blue working solution into each well instead, for a total volume of 25 µL/well.
- Incubate the reaction at room temperature for 20 - 30 minutes, protected from light.
- Add 50 µL of AldeView™ Blue Enhancer (Component C) into each well. For a 384-well plate, add 25 µL of AldeView™ Blue Enhancer into each well.
- Incubate the reaction at room temperature for 20 minutes, protected from light.
- Monitor the absorbance increase with an absorbance plate reader at around 620 to 660 nm (Max at 620 nm).
Citations
Authors: Yang, Guang and Komaki, Yukako and Ibuki, Yuko
Journal: Mutation Research/Genetic Toxicology and Environmental Mutagenesis (2018)
Authors: Kumar, Sudhir and Wang, Jiang and Rani, Richa and G, undefined and hi, Ch and rashekhar R, undefined
Journal: PloS one (2016): e0147864
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
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)
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
Safety data sheet