ALT catalyze the amino conversion reaction between alanine and α-ketoglutaric acid to produce pyruvic acid and glutamic acid at pH 7.4 and 37℃. Then phenylhydrazine was added to form phenylhydrazone with pyruvic acid. Phenylhydrazone is reddish brown under alkaline conditions. ALT activity can be calculated by measuring the OD values at 510 nm.
Performance characteristics
| Synonyms |
ALT,GPT,Glutamic-pyruvic transaminase |
| Sample type |
Serum,plasma,cells,animal tissue |
| Sensitivity |
1.26 IU/L |
| Detection range |
1.26-72.3 IU/L |
| Detection method |
Colorimetric method |
| Assay type |
Enzyme Activity |
| Assay time |
100 min |
| Precision |
Average inter-assay CV: 8.200%Average intra-assay CV: 4.300% |
| Other instruments required |
Micropipettor, Vortex mixer, Incubator |
| Other reagents required |
Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| 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 (1.26-72.3 IU/L).
The recommended dilution factor for different samples is as follows (for reference only):
|
Sample type
|
Dilution factor
|
|
10% Rat liver tissue homogenization
|
30-60
|
|
10% Rat kidney tissue homogenization
|
1
|
|
Human serum
|
1
|
|
Human plasma
|
1
|
|
HepG2 cells homogenization
|
1
|
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
