- Detection method: Colorimetric method
Manual
Detection principle
Acetyl-CoA can react with choline under the catalysis of choline acetyltransferase (ChAT) to produce coenzyme A (CoA), CoA can combine with the 4, 4-dithiopyridine. The activity of ChAT can be calculated indirectly by measuring the OD value at 324 nm.
Performance characteristics
| Synonyms |
ChAT |
| Sample type |
Animal tissue |
| Sensitivity |
1.21 U/g fresh weigh |
| Detection range |
1.21-40 U/g fresh we |
| Detection method |
Colorimetric method |
| Assay type |
Enzyme Activity |
| Assay time |
80 min |
| Precision |
Average inter-assay CV: 9.200%Average intra-assay CV: 5% |
| Other instruments required |
Micropipettor, Centrifuge, Incubator, Water bath, Vortex mixer |
| Other reagents required |
Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| Storage |
Reagent 2, Reagent 3, Reagent 6: -20℃, others: 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 (1.21-40 U/g fresh weight).
The recommended dilution factor for different samples is as follows (for reference only):
|
Sample type
|
Dilution factor
|
|
20% Mouse brain tissue homogenate
|
1-2
|
|
20% Rat heart tissue homogenate
|
1
|
|
20% Rat liver tissue homogenate
|
1
|
|
20% Mouse kidney tissue homogenate
|
1
|
|
20% Rat lung tissue homogenate
|
1
|
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
