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Catalase (CAT) Activity Fluorometric Assay Kit– MSE Supplies LLC

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Catalase (CAT) Activity Fluorometric Assay Kit

SKU: E-BC-F006-500

  • $ 81695



Catalase (CAT) Activity Fluorometric Assay Kit

SKU # E-BC-F006
Detection Instrument Fluorescence microplate reader (Ex/Em=535 nm/587 nm)
Detection Method Fluorescence method


Product Details

Properties

Synonyms CAT
Sample type Serum, plasma, animal tissue
Sensitivity 0.01 U/L
Detection Range 0.01-6.51 U/L
Detection Method Fluorescence method
Assay Time 30 min
Precision Inter-assay CV: 6.8% | intra-assay CV: 3.4%
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
Mouse serum 60-80
Mouse plasma 60-80
Rat serum 70-80
Human saliva 30-50
Rat urine 30-50
10% Mouse liver tissue homogenate 2500-3000
10% Mouse lung tissue homogenate 200-400
10% Rat muscle tissue homogenate 100-200
10% Rat brain tissue homogenate 40-50
10% Mouse kidney tissue homogenate 2000-2500

 

Detection Principle

Catalase can decompose H2O2 to generate H2O and O2, the residual H2O2 in the detection system react with the fluorescent substance, and the content of residual H2O2 is proportional to the fluorescence intensity at the excitation wavelength of 535 nm and emission wavelength of 587 nm, the catalase activity is inversely proportional to the fluorescence intensity.

 

Kit Components & Storage

Item Component Size (96 T) Storage
Reagent 1 Buffer Solution 60 mL ×1 vial 20°C, 12 months
Reagent 2 Substrate 0.1 mL × 1 vial -20°C, 12 months shading light
Reagent 3 Probe Solution 0.12 mL × 1 vial -20°C, 12 months shading light
Reagent 4 Enzyme Reagent Powder × 1 vial -20°C, 12 months shading light
Reagent 5 1 mol/L H2O2 Standard Solution 0.4 mL × 1 vial -20°C, 12 months shading light

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) 0.80 3.00 5.40
    %CV 4.2 3.5 2.5

     

    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) 0.80 3.00 5.40
    %CV 7.5 6.0 6.9

     

    Recovery

    Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 92%.


    Standard 1 Standard 2 Standard 3
    Expected Conc.(μmol/L) 3 25 38
    Observed Conc.(μmol/L) 2.7 23.3 35.3
    Recovery Rate (%) 90 93 93

     

    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 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:

    Concentration (μmol/L) 0 1 5 10 20 30 40 50
    Fluorescence value 102 351 1209 2542 5341 7984 10023 13847
    101 351 1229 2603 5434 8133 10396 13778
    Average fluorescence value 102 351 1219 2572 5387 8058 10209 13812
    Absoluted fluorescence value 0 249 1117 2471 5286 7957 10108 13711