Amplite® Colorimetric Acetylcholinesterase Assay Kit
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12171501 |
Overview | ![]() ![]() |
Platform
Absorbance microplate reader
Absorbance | 410 ± 5 nm |
Recommended plate | Clear bottom |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare AChE working solution (50 µL)
- Add AChE standards or AChE test samples (50 µL)
- Incubate at room temperature for 10 - 30 minutes
- Monitor absorbance at 410 ± 5 nm
Important notes
Thaw all the kit components at room temperature before starting the experiment.
PREPARATION OF STOCK SOLUTION
1. DTNB stock solution (20X):
Add 0.6 mL of Assay Buffer (Component B) into the vial of DTNB (Component A) to make 20X DTNB stock solution. Keep from light. Note: DNTB is not easy to dissolve, it is normal to see the cloudiness of the solution. One can use either the supernatant or the mixture for the experiment.
2. Acetylthiocholine stock solution (20X):
Add 0.6 mL of ddH2O into the vial of Acetylthiocholine (Component C) to make 20X Acetylthiocholine stock solution.
3. Acetylcholinesterase standard solution (50 U/mL):
Add 100 µL of ddH2O with 0.1% BSA into the vial of Acetylcholinesterase Standard (Component D) to make 50 U/mL Acetylcholinesterase standard solution.
PREPARATION OF STANDARD SOLUTION
For convenience, use the Serial Dilution Planner: https://www.aatbio.com/tools/serial-dilution/11400
Add 20 µL of 50 U/mL Acetylcholinesterase standard solution to 980 µL of Assay Buffer (Component B) to generate 1000 mU/mL Acetylcholinesterase standard solution (AS7). Then take 1000 mU/mL Acetylcholinesterase standard solution (AS7) and perform 1:3 serial dilutions in Assay Buffer (Component B) to get serially diluted Acetylcholinesterase standards (AS6 - AS1). Note: Diluted acetylcholinesterase standard solution is unstable and should be used within 4 hours.
PREPARATION OF WORKING SOLUTION
Add 250 μL of 20X DTNB stock solution and 250 μL of 20X Acetylthiocholine stock solution into 4.5 mL of Assay Buffer (Component B) to make a total volume of 5 mL AChE working solution. Keep from light.
For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
SAMPLE EXPERIMENTAL PROTOCOL
Table 1. Layout of Acetylcholinesterase standards and test samples in a white/clear bottom 96-well microplate. AS=Acetylcholinesterase Standards (AS1 - AS7, 1 to 1000 mU/mL); 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 to 1000 mU/mL) |
BL | 50 µL | Assay Buffer (Component B) |
TS | 50 µL | test sample |
- Prepare Acetylcholinesterase 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: Treat cells or tissue samples as desired.
- Add 50 µL of AChE working solution to each well of Acetylcholinesterase standard, blank control, and test samples to make the total Acetylcholinesterase assay volume of 100 µL/well. For a 384-well plate, add 25 µL of AChE working solution into each well instead, for a total volume of 50 µL/well.
- Incubate the reaction for 10 to 30 minutes at room temperature, protected from light.
- Monitor the absorbance increase with an absorbance microplate reader at 410 ± 5 nm.
Images






Citations
Authors: Desprez, Charlotte and Danovi, Davide and Knowles, Charles H and Day, Richard M
Journal: Journal of Tissue Engineering (2023): 20417314221139794
Authors: Jamadagni, Shrirang B and Ghadge, Pooja M and Tambe, Mukul S and Srinivasan, Marimuthu and Prasad, Goli Penchala and Jamadagni, Pallavi S and Prasad, Shyam Baboo and Pawar, Sharad D and Gurav, Arun M and Gaidhani, Sudesh N and others,
Journal: Pharmacognosy Magazine (2023): 09731296221145063
Authors: Xu, Tuan and Li, Shuaizhang and Li, Andrew J and Zhao, Jinghua and Sakamuru, Srilatha and Huang, Wenwei and Xia, Menghang and Huang, Ruili
Journal: Journal of Chemical Information and Modeling (2023)
Authors: Shaban, Nadia Z and Abd El-Kader, Sara E and Mogahed, Fayed AK and El-Kersh, Mohamed AL and Habashy, Noha H
Journal: Scientific Reports (2021): 1--18
Authors: Tsay, Gregory J and Lin, Yu-Ta and Hsu, Chia-Hong and Tang, Feng-Yao and Kuo, Yueh-Hsiung and Chao, Che-Yi
Journal: Biochemistry and Biophysics Reports (2021): 101020
Authors: Li, Shuaizhang and Li, Andrew J and Travers, Jameson and Xu, Tuan and Sakamuru, Srilatha and Klumpp-Thomas, Carleen and Huang, Ruili and Xia, Menghang
Journal: SLAS DISCOVERY: Advancing the Science of Drug Discovery (2021): 24725552211030897
Authors: Chandran, Remya and Vijayan, Dileep and Reddy, Eeda Koti and Kumar, Mantosh and Kesavan, Lakshmi and Jacob, Reena and Ayyiliyath, Sajith and Variyar, Jayadevi and Anwar, Shaik and Zhang, Kam and others,
Journal: bioRxiv (2021)
Authors: Lim, Gyeong Taek and You, Dong Gil and Han, Hwa Seung and Lee, Hansang and Shin, Sol and Oh, Byeong Hoon and Kumar, EK Pramod and Um, Wooram and Kim, Chan Ho and Han, Seungsu and others,
Journal: Journal of extracellular vesicles (2021): e12077
Authors: Li, Shuaizhang and Zhao, Jinghua and Huang, Ruili and Travers, Jameson and Klumpp-Thomas, Carleen and Yu, Wenbo and MacKerell Jr, Alexander D and Sakamuru, Srilatha and Ooka, Masato and Xue, Fengtian and others,
Journal: Environmental health perspectives (2021): 047008
Authors: Ishimota, Makoto and Tajiki-Nishino, Risako and Fukuyama, Tomoki and Tomiyama, Naruto and Sakamoto, Masaki and Ohyama, Kazutoshi
Journal: Water Air Soil Pollut (2020): 287
References
Authors: Seki M, Kosai K, Hara A, Imamura Y, Nakamura S, Kurihara S, Izumikawa K, Kakeya H, Yamamoto Y, Yanagihara K, Miyazaki Y, Mukae H, Tashiro T, Kohno S.
Journal: Jpn J Infect Dis (2009): 6
Authors: Artim DE, Kullmann FA, Daugherty SL, Wu HY, de Groat WC.
Journal: Am J Physiol Renal Physiol (2009): F333
Authors: Green KD, Fridman M, Garneau-Tsodikova S.
Journal: Chembiochem (2009): 2191
Authors: Sarma K, Pal AK, Sahu NP, Ayyappan S, Baruah K.
Journal: Sci Total Environ (2009): 3668
Authors: Castro DP, Figueiredo MB, Genta FA, Ribeiro IM, Tomassini TC, Azambuja P, Garcia ES.
Journal: J Insect Physiol (2009): 532
Authors: Bihaqi SW, Sharma M, Singh AP, Tiwari M.
Journal: J Ethnopharmacol (2009): 409
Authors: Inoue R, Jensen LJ, Jian Z, Shi J, Hai L, Lurie AI, Henriksen FH, Salomonsson M, Morita H, Kawarabayashi Y, Mori M, Mori Y, Ito Y.
Journal: Circ Res (2009): 1399
Authors: Detopoulou P, Nomikos T, Fragopoulou E, Antonopoulou S, Kotroyiannis I, Vassiliadou C, Panagiotakos DB, Chrysohoou C, Pitsavos C, Stefanadis C.
Journal: Clin Biochem (2009): 44
Authors: Ruan CJ, Si JY, Zhang L, Chen DH, Du GH, Su L.
Journal: Neurosci Lett (2009): 159
Authors: Tsoukatos DC, Brocheriou I, Moussis V, Panopoulou CP, Christofidou ED, Koussissis S, Sismanidis S, Ninio E, Siminelakis S.
Journal: J Lipid Res (2008): 2240
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