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Acetylcholinesterase (AchE) Activity Assay Kit– MSE Supplies LLC

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Acetylcholinesterase (AchE) Activity Assay Kit

SKU: E-BC-K174-M-96

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Acetylcholinesterase (AchE) Activity Assay Kit

SKU # E-BC-K174-M
Detection Instrument Microplate reader (optimum wavelength: 412 nm)
Detection method Colorimetric method


Product Details

Properties

Synonyms AchE
Sample Type Serum, plasma, animal tissue and cell
Sensitivity 1.225 U/mL
Detection Range 1.225-490 U/mL
Detection Method Colorimetric method
Assay type Enzyme Activity
Assay time 20 min
Precision Average inter-assay CV: 9.300% | Average intra-assay CV: 4.700%
Other instruments required Micropipettor, Incubator, Vortex mixer, Centrifuge
Other reagents required Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4)
Storage 2-8℃
Valid period 12 months


Images

A F Odetayo et al investigate the effect of bisphenol F on sexual performance and quality. Acetylcholinesterase (AchE) activity of rat plasma was determined using AchE activity assay kit (E-BC-K174-M).

The activity of AchE was significantly decreased comparing with control groups. (P<0.05 versus age-matched control)

 

Dilution of Sample

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (13.88-320.13 U/mL).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type Dilution factor
Mouse serum 8-20
Mouse plasma 4-10
Human serum 4-10
Human plasma 4-10
Rat serum 4-10
Dog serum 4-10
Horse serum 2-8
10% Mouse liver tissue homogenate 1
10% Mouse kidney tissue homogenate 1
10% Mouse brain tissue homogenate 2-8
10% Crucian carp muscle tissue homogenate 1

 

Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).

 

Detection Principle

AchE catalyzes the hydrolysis of acetylcholine to form choline, and choline react with dithio p-nitrobenzoic acid (DTNB) to form 5-mercapto-nitrobenzoic acid (TNB). TNB has an absorption peak at 412nm. And the activity of AchE is calculated by measuring the increasing rate of absorbance at 412nm.

 

Kit Components & Storage

Item Component Size 1(48 T) Size 2(96 T) Storage
Reagent 1 Lysis Buffer 50 mL × 1 vial 50 mL × 2 vials 2-8°C, 12 months
Reagent 2 Buffer Solution 15 mL × 1 vial 30 mL × 1 vial 2-8°C, 12 months
Reagent 3 Chromogenic Agent Powder × 1 vial Powder × 1 vial 2-8°C, 12 months,
shading light
Reagent 4 Substrate Powder × 1 vial Powder × 1 vial 2-8°C, 12 months,
shading light
Microplate 96 wells No requirement
Plate Sealer 2 pieces

 

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.

 

Technical Data:

Parameter:

Intra-assay Precision

Three human serum samples were assayed in replicates of 20 to determine precision within an assay. (CV = Coefficient of Variation)

Parameters Sample 1 Sample 2 Sample 3
Mean (U/mL) 10.20 88.50 264.00
%CV 5.2 4.6 4.3


Inter-assay Precision

Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Parameters Sample 1 Sample 2 Sample 3
Mean (μg/mL) 10.20 88.50 264.00
%CV 9.1 9.3 9.5

 

Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 104%.

Standard 1 Standard 2 Standard 3
Expected Conc. (U/mL) 138.5 252 381.3
Observed Conc. (U/mL) 146.8 257.0 396.6
Recovery rate (%) 106 102 104

 

Sensitivity

The analytical sensitivity of the assay is 1.225 U/mL. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.