Test Principle
The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Human ALT. Standards or samples are added to the appropriate microtiter plate wells then with a biotin-conjugated antibody specific to Human ALT. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Human ALT, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of Human ALT in the samples is then determined by comparing the OD of the samples to the standard curve.
Product Specification
| Product Name | Human ALT(Alanine Aminotransferase) ELISA Kit |
| Alias | GPT; ALAT; SGPT; AAT1; GPT1; Serum Glutamic Pyruvic Transaminase; Alanine Transaminase; Glutamate pyruvate transaminase 1; Glutamic--alanine transaminase 1 |
| UniProt ID | P24298 |
| Sample Type | serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids |
| Assay Type | Sandwich |
| Research Area | Enzyme & Kinase;Infection immunity;Hepatology |
| Assay length | 3.5h |
Standard Curve
| Concentration (U/L) | OD | Corrected OD |
| 10.00 | 1.964 | 1.870 |
| 5.00 | 1.616 | 1.522 |
| 2.50 | 1.043 | 0.949 |
| 1.25 | 0.769 | 0.675 |
| 0.63 | 0.541 | 0.447 |
| 0.32 | 0.319 | 0.225 |
| 0.16 | 0.256 | 0.162 |
| 0.00 | 0.094 | 0.000 |
Precision
- Intra-assay Precision (Precision within an assay)：CV%<8%
- Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
- Inter-assay Precision (Precision between assays)：CV%<10%
- Three samples of known concentration were tested in forty separate assays to assess inter-assay precision.
Recovery
Matrices listed below were spiked with certain level of recombinant ALT and the recovery rates were calculated by comparing the measured value to the expected amount of ALT in samples
| Matrix | Recovery Range(%) | Average Recovery(%) |
| serum(n=5) | 83-95% | 89% |
| EDTA plasma(n=5) | 88-102% | 95% |
| Heparin plasma(n=5) | 79-97% | 88% |
Linearity
| Matrix | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 86-99% | 93-107% | 85-94% | 87-101% |
| EDTA plasma(n=5) | 86-97% | 88-95% | 92-101% | 87-98% |
| Heparin plasma(n=5) | 85-92% | 79-96% | 89-100% | 85-96% |
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