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Alanine Aminotransferase (ALT/GPT) Activity Fluorometric Assay Kit– MSE Supplies LLC

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Alanine Aminotransferase (ALT/GPT) Activity Fluorometric Assay Kit - MSE Supplies LLC

Alanine Aminotransferase (ALT/GPT) Activity Fluorometric Assay Kit

SKU: E-BC-F038-96

  • £27600
  • Save £4800

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