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Na+K (Sodium Potassium)+-ATPase Activity Assay Kit– MSE Supplies LLC

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Na+K (Sodium Potassium)+-ATPase Activity Assay Kit

SKU: E-BC-K539-M-96

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Na+K (Sodium Potassium)+-ATPase Activity Assay Kit

SKU # E-BC-K539-M
Detection Instrument Microplate reader (640 nm-680 nm, optimum wavelength: 660 nm)
Detection method Colorimetric method


Product Details

Properties

Synonyms Na+K+-ATPase
Sample type Serum, plasma, animal tissue
Sensitivity 0.11 μmol Pi/mL/hour
Detection range 0.42-4.99 μmol Pi/mL
Detection Method Colorimetric method
Assay type Enzyme Activity
Assay time 35 min
Precision Average inter-assay CV: 2.200% | Average intra-assay CV: 2.200%
Other instruments required Micropipettor, Tubes, Vortex mixer, Incubator, 37℃ Water bath, Centrifuge.
Other reagents required Normal saline (0.9% NaCl)
Storage 2-8℃
Valid period 12 months


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 ( 0.42-4.99 μmol Pi/mL/hour).

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

Sample type Dilution factor
Rat plasma 1
10% Rat liver tissue homogenate 6-10
10% Rat spleen tissue homogenate 4-8
10% Rat heart tissue homogenate 4-8
10% Mouse brain tissue homogenate 4-8

 

Note: The diluent is normal saline (0.9% NaCl).

 

Detection Principle

Na+K+-ATPase decomposes ATP to produce ADP and Phosphorus, and calculates the activity of Na+K+-ATPase by measuring the content of phosphorus.

 

Kit Components & Storage

Item Component Size (96 T) Storage
Reagent 1 Buffer Solution 6 mL × 1 vial 2-8°C, 12 months
Reagent 2 Accelerant A 5 mL × 1 vial 2-8°C, 12 months
Reagent 3 Substrate Powder × 1 vial 2-8°C, 12 months
Reagent 4 Accelerant B 1.5 mL × 1 vial 2-8°C, 12 months
Reagent 5 Protein Precipitator 1.5 mL × 2 vials 2-8°C, 12 months
Reagent 6 Powder A Powder × 1 vial 2-8°C, 12 months,
shading light
Reagent 7 Powder B Powder × 1 vial 2-8°C, 12 months,
shading light
Reagent 8 Acid Reagent 5 mL × 1 vial 2-8°C, 12 months
Reagent 9 10 μmol/mL Pi Standard 2 mL × 1 vial 2-8°C, 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 (μmol Pi/mL/hour) 0.85 1.35 2.80
%CV 2.5 2.2 1.9


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 (μmol Pi/mL/hour) 0.85 1.35 2.80
%CV 1.9 2.4 2.3


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 104%.


Standard 1 Standard 2 Standard 3
Expected Conc. (μmol/L) 0.35 1.05 2.3
Observed Conc. (μmol/L) 0.4 1.1 2.4
Recovery rate (%) 101 106 105

 

Sensitivity

The analytical sensitivity of the assay is 0.11 μmol Pi/mL/hour. 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 0.25 0.5 1.0 1.5 2.0 2.5 3.0
OD value 0.037 0.161 0.283 0.522 0.766 1.021 1.259 1.535
0.038 0.161 0.283 0.520 0.765 1.004 1.255 1.517
Average OD 0.038 0.161 0.283 0.521 0.766 1.012 1.257 1.526
Absoluted OD 0 0.123 0.245 0.483 0.728 0.974 1.219 1.488