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Cholinesterase (CHE) Activity Assay Kit
SKU: E-BC-K052-S-100
Cholinesterase (CHE) Activity Assay Kit
| SKU # | E-BC-K052-S |
| Detection Instrument | Spectrophotometer(520 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | CHE |
| Sample Type | Whole blood, serum, plasma, tissue, cells |
| Sensitivity | 1.17 U/mL |
| Detection Range | 1.17-160 U/mL |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 60 min |
| Precision | Average inter-assay CV: 9.400% | Average intra-assay CV: 3.700% |
| Other instruments required | Micropipettor, Centrifuge, Incubator, Water bath, Vortex mixer |
| Other reagents required | Normal saline (0.9% NaCl) |
| Storage | 2-8℃ |
| Valid period | 12 months |
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 (1.17-160 U/mL).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 2-3 |
| Human plasma | 2-3 |
| Mouse serum | 2-3 |
| Mouse plasma | 2-3 |
| SH-SY5Ycells | 1 |
| 10% Mouse brain tissue homogenate | 1 |
| 10% Rat brain tissue homogenate | 1 |
| 10% Rat kidney tissue homogenate | 1 |
| 10% Rat spleen tissue homogenate | 1 |
| 10% Mouse liver tissue homogenate | 1 |
| 10% Mouse brain tissue homogenate | 1 |
Note: The diluent is normal saline (0.9% NaCl).
Detection Principle
Cholinesterase breaks down acetylcholine into choline and acetic acid. Acetylcholine that is not hydrolyzed by cholinesterase reacts with basic hydroxylamine to form acetamidamine. It reacts in an acidic solution to form a brown-red hydroxamate iron complex. The color depth is directly proportional to the amount of remaining acetylcholine, which can be colorimetrically quantified. Cholinesterase activity was calculated.
Kit Components & Storage
| Item | Component | Size 1 (50 assays) | Size 2 (100 assays) | Storage |
| Reagent 1 | Buffer Solution | 60 mL × 1 vial | 60 mL × 2 vials | 2-8℃, 12 months |
| Reagent 2 | Substrate | Powder × 1 vial | Powder × 2 vials | 2-8°C, 12 months, shading light |
| Reagent 3 | Diluent 1 | 5 mL ×1 vial | 10 mL ×1 vial | 2-8°C, 12 months |
| Reagent 4 | Chromogenic Agent 1 | Powder × 1 vial | Powder × 1 vial | 2-8℃, 12 months |
| Reagent 5 | Alkali Reagent | 30 mL ×1 vial | 60 mL × 1 vial | 2-8℃, 12 months |
| Reagent 6 | Acid Reagent | 30 mL × 1 vial | 60 mL × 1 vial | 2-8°C, 12 months |
| Reagent 7 | Protein Precipitator | 20 mL × 1 vial | 40 mL × 1 vial | 2-8°C, 12 months |
| Reagent 8 | Chromogenic Agent 2 | Powder × 1 vial | Powder × 1 vial | 2-8°C, 12 months, shading light |
| Reagent 9 | Diluent 2 | 1 mL × 1 vial | 2 mL × 1 vial | 2-8°C, 12 months |
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) | 2.60 | 46.50 | 133.00 |
| %CV | 4.2 | 3.6 | 3.3 |
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 (U/mL) | 2.60 | 46.50 | 133.00 |
| %CV | 9.5 | 9.1 | 9.6 |
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) | 15.6 | 66.7 | 142.5 |
| Observed Conc. (U/mL) | 15.9 | 70.7 | 148.2 |
| Recovery rate (%) | 102 | 106 | 104 |
Sensitivity
The analytical sensitivity of the assay is 1.17 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