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Phosphofructokinase (PFK) Activity Assay Kit– MSE Supplies LLC

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Phosphofructokinase (PFK) Activity Assay Kit

SKU: E-BC-K612-M-96

  • $ 39295
  • Save $ 7000



Phosphofructokinase (PFK) Activity Assay Kit

SKU # E-BC-K612-M
Detection Instrument Microplate reader (330-350 nm, optimum wavelength: 340 nm)
Detection method Colorimetric method


Product Details

Properties

Synonyms PFK
Sample type Serum, plasma, tissue and cell sample
Sensitivity 0.27 U/L
Detection range 0.27-32.29 U/L
Detection Method Colorimetric method
Assay type Enzyme Activity
Assay time 15 min
Precision Average inter-assay CV: 8% | Average intra-assay CV: 3%
Other instruments required Centrifuge
Other reagents required Normal saline (0.9% NaCl)
Storage -20℃
Valid period 12 months


Dilution of Sample

The recommended dilution factor for different samples is as follows (for reference only):

Sample type Dilution factor
10% Rat liver tissue homogenate 1
10% Rat kidney tissue homogenate 1
10% Rat brain tissue homogenate 1
10% Mouse liver tissue homogenate 1
10% Mouse spleen tissue homogenate 1
10% Mouse kidney tissue homogenate 1
10% Mouse heart tissue homogenate 1
Rat serum 1
Rat plasma 1
Jurkat cell 1

 

Note: The diluent is normal saline (0.9% NaCl). For the dilution of other sample types, please do pretest to confirm the dilution factor.

 

Detection Principle

Phosphofructokinase(also known as 6-phosphofructokinase; PFK) is a class of kinases that act on fructose-6-phosphate. Phosphofructokinase catalyzes fructose-6-phosphate and ATP to produce fructose-1, 6-diphosphate and ADP, and pyruvate kinase and lactate dehydrogenase further catalyze the oxidation of NADH to NAD+. The activity of PFK can be reflected by the determination of NADH decline rate at 340 nm.

 

Kit Components & Storage

Item Component Size 1(48 T) Size 2(96 T) Storage
Reagent 1 Buffer Solution 10 mL ×1 vial 20 mL × 1 vial -20℃, 12 months,
shading light
Reagent 2 Substrate A Powder × 1 vial Powder × 2 vials -20°C, 12 months shading light
Reagent 3 Substrate B  Powder × 1 vial Powder × 2 vials -20°C, 12 months shading light
Reagent 4 Enzyme Reagent Powder × 1 vial Powder × 2 vials -20°C, 12 months shading light
UV 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) 2.60 18.40 26.50
%CV 3.5 3.1 2.4


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) 2.60 18.40 26.50
%CV 7.6 8.5 7.9


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 100%


Sample 1 Sample 2 Sample 3
Expected Conc. (U/L) 8.5 22.3 29.5
Observed Conc. (U/L) 8.4 22.7 29.2
Recovery rate (%) 99 102 99

 

Sensitivity

The analytical sensitivity of the assay is 0.27 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.