Mak 119 Bul
Mak 119 Bul
Mak 119 Bul
The Acetylcholinesterase Activity Assay kit provides a Reagents and Equipment Required but Not
simple and direct procedure for measuring AChE levels Provided.
in a variety of samples such as blood, serum, and Spectrophotometric multiwell plate reader
plasma. This assay is an optimized version of the Clear 96 well flat-bottom plate
Ellman method in which thiocholine, produced by
AChE, reacts with 5,5-dithiobis(2-nitrobenzoic acid) to Precautions and Disclaimer
form an colorimetric (412 nm) product, proportional to This product is for R&D use only, not for drug,
the AChE activity present. One unit of AChE is the household, or other uses. Please consult the Material
amount of enzyme that catalyzes the production of Safety Data Sheet for information regarding hazards
1.0 mole of thiocholine per minute at pH 7.5 at room and safe handling practices.
temperature. This kit has a linear range of
10600 units/L of AChE activity. Storage/Stability
This kit is shipped, and should be stored at room
temperature.
Procedure
Sample Preparation
Blood samples should be diluted 40-fold in Assay
Buffer.
4. Incubate the samples at room temperature. After One unit of AChE is the amount of enzyme that
2 minutes, take the initial absorbance measurement catalyzes the production of 1.0 mole of thiocholine per
at 412 nm (A412)initial. minute at room temperature at pH 7.5. This kit has a
linear range of 10600 units/L of AChE activity.
5. Continue to incubate the plate at room temperature.
At 10 minutes, take the final measurement (A412)final.
Troubleshooting Guide
Problem Possible Cause Suggested Solution
Omission of step in procedure Refer and follow Technical Bulletin precisely
Plate reader at incorrect wavelength Check filter settings of instrument
Assay not working
For UV assays, use clear plates that are UV
Type of 96 well plate used
transparent or quartz plates.
Incorrect volumes used Use calibrated pipettes and aliquot correctly
Samples with erratic
Samples measured at incorrect
readings Check the equipment and filter settings
wavelength
LS,MAM 11/13-1
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