- Detection method: Colorimetric method
Manual
Detection principle
AST enables alpha-ketoglutaric acid and aspartic acid to displace amino to form glutamic acid and oxaloacetic acid. Oxaloacetic acid can decarboxylate itself to form Pyroracemic acid during the reaction. Pyroracemic acid reacted with 2,4-dinitrophenylhydrazine (DNPH) to form 2,4-dinitrophenylhydrazone showing reddish brown in alkaline solution. Measure the OD values and calculate the enzyme activity.
Performance characteristics
| Synonyms |
AST,GOT |
| Sample type |
Serum, plasma, animal tissue |
| Sensitivity |
0.38 IU/L |
| Detection range |
0.38-72.30 IU/L |
| Detection method |
Colorimetric method |
| Assay type |
Enzyme Activity |
| Assay time |
100 min |
| Precision |
Average inter-assay CV: 4.900%Average intra-assay CV: 4.200% |
| Other instruments required |
Test tube, Micropipettor, 37℃ 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 (0.38-72.30 IU/L).
The recommended dilution factor for different samples is as follows (for reference only):
|
Sample type
|
Dilution factor
|
|
Human plasma
|
1
|
|
Human serum
|
1
|
|
10% Rat liver tissue homogenate
|
16-32
|
|
HepG2 supernatant
|
1
|
|
Rat serum
|
1
|
|
Mouse plasma
|
1
|
|
10% Rat kidney tissue homogenate
|
1
|
|
10% Rat heart tissue homogenate
|
1
|
Note: The diluent is normal saline (0.9% NaCl).
