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Alanine Aminotransferase (ALT/GPT) Activity Fluorometric Assay Kit
SKU: E-BC-F038-96
Alanine Aminotransferase (ALT/GPT) Activity Fluorometric Assay Kit
| SKU # | E-BC-F038 |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=535 nm/587 nm) |
| Detection Method | Fluorescence method |
Product Details
Properties
| Synonyms | ALT |
| Sample Type | Serum, plasma, animal tissue |
| Sensitivity | 0.01 U/L |
| Detection Range | 0.01-0.83 U/L |
| Detection Method | Fluorescence method |
| Assay Time | 75 min |
| Precision | Inter-assay CV: 9.8% | intra-assay CV: 2.3% |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Valid period | 12 months |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
|---|---|
| Human serum | 10-15 |
| Dog serum | 5-10 |
| Rat serum | 10-15 |
| 10% Mouse heart tissue homogenate | 100-120 |
| 10% Rat spleen tissue homogenate | 10-15 |
| 10% Rat liver tissue homogenate | 300-500 |
| 10% Rat kidney tissue homogenate | 100-120 |
| 10% Rat lung tissue homogenate | 100-120 |
The diluent is buffer solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
ALT catalyze the amino conversion reaction between alanine and α-ketoglutaric acid to produce pyruvic acid and glutamic acid. Under the action of pyruvate oxidase, pyruvic acid generates H2O2, which reacts with the non-fluorescent substance to form fluorescent substance under the action of peroxidase. The activity of ALT can be calculated by measuring the increase of fluorescence value at the excitation wavelength of 535 nm and the emission wavelength of 590 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 60 mL ×1 vial | 60 mL ×2 vials | -20℃, 12 months |
| Reagent 2 | Probe Solution | 0.25 mL × 1 vial | 0.5 mL × 1 vial | -20℃, 12 months shading light |
| Reagent 3 | Enzyme Reagent | Powder × 1 vial | Powder × 2 vials | -20℃, 12 months |
| Reagent 4 | Substrate Solution | 1.2 mL ×1 vial | 1.2 mL × 2 vials | -20℃, 12 months shading light |
| Reagent 5 | 100 mmol/L Pyruvate Standard |
0.1 mL ×1 vial | 0.1 mL × 1 vial | -20℃, 12 months |
| Black Microplate | 96 wells | No requirement | ||
| Plate Sealer | 2 pieces | |||
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/L) | 0.10 | 0.35 | 0.65 |
| %CV | 2.5 | 2.3 | 2.1 |
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/L) | 0.10 | 0.35 | 0.65 |
| %CV | 9.9 | 10.0 | 9.5 |
Sensitivity
The analytical sensitivity of the assay is 0.01 U/L. 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.
Standard Curve:
As the fluorescence value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:
| Concentration (μmol/L) | 0 | 10 | 20 | 25 | 30 | 35 | 40 | 50 |
| Fluorescence value | 1087 | 1806 | 2567 | 2922 | 3379 | 3906 | 4336 | 5384 |
| 1150 | 1804 | 2550 | 2942 | 3362 | 3862 | 4408 | 5415 | |
| Average fluorescence value | 1118 | 1805 | 2559 | 2932 | 3370 | 3884 | 4372 | 5399 |
| Absoluted fluorescence value | 0 | 687 | 1440 | 1814 | 2252 | 2766 | 3254 | 5399 |