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Alanine Aminotransferase (ALT/GPT) Activity Assay Kit
SKU: E-BC-K235-M-500
Alanine Aminotransferase (ALT/GPT) Activity Assay Kit
| SKU # | E-BC-K235-M |
| Detection Instrument | Microplate reader (500-520 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | ALT, GPT, Glutamic-pyruvic transaminase |
| Sample Type | Serum, plasma, animal tissue, cells, cell culture supernatant |
| Sensitivity | 0.75 IU/L |
| Detection Range | 0.75-72.3 IU/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 80 min |
| Precision | Average inter-assay CV: 9% | Average intra-assay CV: 5.300% |
| Other instruments required | Micropipettor, Vortex mixer, Incubator, Multichannel pipette |
| Other reagents required | Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Images
X Zhang et al investigate the usage of H2O2 and O2 for cooperative chemodynamic/ starvation cancer therapy. Alanine aminotransferase (ALT) activity in mouse blood was determined using ALT activity assay kit (E-BC-K235-M).
No significant change of ALT activity was observed, the LCFG nanoparticles did not include hepatotoxicity.
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 (0.75-72.3 IU/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Human plasma | 1 |
| Porcine serum | 1 |
| Rat serum | 1 |
| 10% Rat brain tissue homogenization | 1 |
| 10% Rat heart tissue homogenization | 1 |
| 10% Rat liver tissue homogenization | 40-60 |
| 10% Rat kidney tissue homogenization | 1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
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.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 0.5 mL × 1 vial | 0.5 mL × 1 vial | 2-8℃, 12 months |
| Reagent 2 | 2 mmol/L Sodium Pyruvate |
0.5 mL × 1 vial | 0.5 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | Substrate Solution | 2.5 mL × 1 vial | 5 mL × 1 vial | 2-8℃, 12 months |
| Reagent 4 | Chromogenic Agent | 2.5 mL × 1 vial | 5 mL × 1 vial | 2-8°C, 12 months, shading light |
| Reagent 5 | Alkali Reagent | 2.5 mL × 1 vial | 5 mL × 1 vial | 2-8℃, 12 months |
| 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 (IU/L) | 3.50 | 26.50 | 52.60 |
| %CV | 5.7 | 5.2 | 5.0 |
Inter-assay Precision
Three human serum samples were assayed 17 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (IU/L) | 3.50 | 26.50 | 52.60 |
| %CV | 8.4 | 9.2 | 9.4 |
Sensitivity
The analytical sensitivity of the assay is 0.75 IU/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 OD 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:
| Carmen Unit | 0 | 28 | 57 | 97 | 150 | 200 |
| Average OD | 0.247 | 0.324 | 0.384 | 0.436 | 0.489 | 0.537 |
| Absoluted OD | 0 | 0.078 | 0.137 | 0.190 | 0.242 | 0.290 |