Amplite® Fluorimetric Butyrylcholinesterase Activity Assay Kit
Price | |
Catalog Number | |
Unit Size | |
Quantity |
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 |
Intended use | Research Use Only (RUO) |
Overview | SDSProtocol |
Platform
Fluorescence microplate reader
Excitation | 490 nm |
Emission | 525 nm |
Cutoff | 515 nm |
Recommended plate | Solid black |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare BChE standards, dye working solution and test samples
- Add BChE standards or test samples (100 uL)
- Add BChE dye working solution (100 uL)
- Incubate at room temperature for 10-30 minutes
- Monitor fluorescence intensity at Ex/Em=490/525 nm
Important notes
Thaw all the kit components at room temperature before starting the experiment.
PREPARATION OF STOCK SOLUTION
- Thiolite™ Green stock solution (200X):
Add 50 uL of DMSO (Component E) into the vial of Thiolite™ Green (Component A) to make 200X Thiolite™ Green stock solution.
- Butyrylthiocholine (BTC) stock solution (100X):
Add 120 uL of ddH2O into the vial of BTC (Component C) to make 100X BTC stock solution.
- Butyrylcholinesterase (BChE) Standard solution (5 U/mL):
Add 50 uL of ddH2O with 0.1% BSA into the vial of BChE Standard (Component D) to make 5 U/mL BChE standard solution.
PREPARATION OF STANDARD SOLUTION
For convenience, use the Serial Dilution Planner: https://www.aatbio.com/tools/serial-dilution/11407
Add 20 uL of 5 U/mL BChE standard solution to 980 uL of Assay Buffer (Component B) to generate 100 mU/mL BChE standard solution (BS1). Then take 100 mU/mL BChE standard solution (BS1) and perform 1:3 serial dilutions in Assay Buffer(Component B) to get serially diluted BChE standards (BS2 - BS7). Note: Diluted BChE standard solution is unstable and should be used within 4 hours.
PREPARATION OF WORKING SOLUTION
BChE dye working solution:
Add 50 uL of 200X Thiolite™ Green stock solution and 100 uL of 100X BTC stock solution into 10 mL of Assay Buffer (Component B) to make a total volume of 10.1 mL BChE dye working solution. Note: Keep it from light.
SAMPLE EXPERIMENTAL PROTOCOL
Table 1. Layout of BChE standards and test samples in a clear bottom 96-well microplate. BS=BChE standards (BS1-BS7, 100 to 0.13 mU/mL); BL=Blank Control; TS=Test Samples
BL |
BL |
TS |
TS |
BS1 |
BS1 |
… |
… |
BS2 |
BS2 |
… |
… |
BS3 |
BS3 |
|
|
BS4 |
BS4 |
|
|
BS5 |
BS5 |
|
|
BS6 |
BS6 |
|
|
BS7 |
BS7 |
|
|
Table 2. Reagent composition for each well.
Well |
Volume |
Reagent |
BS1-BS7 |
100 uL |
Serial Dilutions (100 to 0.13 mU/mL) |
BL |
100 uL |
Assay Buffer (Component B) |
TS |
100 uL |
Test Sample |
- Prepare BChE standards (BS), blank controls (BL), and test samples (TS) according to the layout provided in Tables 1 and 2. For a 384-well plate, use 25 uL of reagent per well instead of 100 uL. Note: Treat cells or tissue samples as desired.
- Add 100 uL of BChE dye working solution to each well of BChE standard, blank control, and test samples to make the total assay volume 200 uL/well. For a 384-well plate, add 25 uL of BChE working solution into each well instead, for a total volume of 50 uL/well.
- Incubate the reaction for 10 to 30 minutes at room temperature, protected from light.
- Monitor the fluorescence increase with an fluorescence microplate reader at Ex/Em=490/525 nm.
Product Family
Name | Excitation (nm) | Emission (nm) |
Amplite® Fluorimetric HDAC Activity Assay Kit *Green Fluorescence* | 493 | 520 |
Images
Citations
Authors: Chen, G., Feng, H., Xi, W., Xu, J., Pan, S., Qian, Z.
Journal: Analyst (2019): 559-566
Authors: Orhan, I. E., Senol Deniz, F. S., Traedal-Henden, S., Ceron-Carrasco, J. P., den Haan, H., Pena-Garcia, J., Perez-Sanchez, H., Emerce, E., Skalicka-Wozniak, K.
Journal: Chem Biodivers (2019): e1900017
Authors: Jasiecki, J., Zuk, M., Krawczynska, N., Jonca, J., Szczoczarz, A., Lew and owski, K., Waleron, K., Wasag, B.
Journal: Chem Biol Interact (2019): 154-157
Authors: Jiang, C. S., Ge, Y. X., Cheng, Z. Q., Wang, Y. Y., Tao, H. R., Zhu, K., Zhang, H.
Journal: Molecules (2019): se name="11406.enl" path="C:\Users\aatbi\Drop
Authors: Maximova, N., Caddeo, G., Zanon, D., Maestro, A., Simeone, R.
Journal: J Clin Med (2019): se name="11406.enl" path="C:\Users\aatbi\Drop
Authors: Mughal, E. U., Sadiq, A., Ashraf, J., Zafar, M. N., Sumrra, S. H., Tariq, R., Mumtaz, A., Javid, A., Khan, B. A., Ali, A., Javed, C. O.
Journal: Bioorg Chem (2019): 103124
Authors: Gao, X. H., Tang, J. J., Liu, H. R., Liu, L. B., Liu, Y. Z.
Journal: Drug Dev Res (2019): 438-445
Authors: Bursal, E., Aras, A., Kilic, O., Taslimi, P., Goren, A. C., Gulcin, I.
Journal: J Food Biochem (2019): e12776
Authors: Vuksic, A., Lovric, J., Konjevoda, P., Blazevic, N., Bilusic, M., Bradamante, V.
Journal: Arh Hig Rada Toksikol (2019): 30-35
Authors: Dorling, J. L., Clayton, D. J., Jones, J., Carter, W. G., Thackray, A. E., King, J. A., Pucci, A., Batterham, R. L., Stensel, D. J.
Journal: Am J Clin Nutr (2019): ersion="1.0" encoding="UTF-8" ?>11406.enlEndN
Application notes
Acetylcholinesterase Inhibitory Activity of Pigment Echinochrome A
Ameliorative Effect of Novel Vitamin Formula with Herbal Extracts on Scopolamine-Induced Alzheimer's Disease
An Increase in Plasma Homovanillic Acid with Cocoa Extract Consumption Is Associated with the Alleviation of Depressive Symptoms in Overweight or Obese Adults
Attenuation of lysyl oxidase and collagen gene expression in keratoconus patient corneal epithelium corresponds to disease severity