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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 |