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Ca2+-ATPase Activity Assay Kit
SKU: E-BC-K212-M-96
Ca2+-ATPase Activity Assay Kit
| SKU # | E-BC-K212-M |
| Detection Instrument | Microplate reader (660 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | Ca2+-ATPase |
| Sample Type | Animal tissue |
| Sensitivity | 1.18 U/kg wet weight |
| Detection Range | 1.18-286.43 U/kg wet weight |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 90 min |
| Precision | Average inter-assay CV: 8% | Average intra-assay CV: 6% |
| Other instruments required | Test tube, Vortex mixer, Incubator, Centrifuge, 100°C water bath, Microplate reader |
| Other reagents required | Ultrapure water, Normal saline (0.9% NaCl) |
| Storage | 2-8℃ |
| 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 | 5-8 |
| 10% Mouse liver tissue homogenate | 1 |
| 10% Rat heart tissue homogenate | 5-8 |
| 10% Rat lung tissue homogenate | 5-8 |
| 10% Rat kidney tissue homogenate | 5-8 |
| 10% Rat brain tissue homogenate | 2-3 |
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
ATPase can decompose ATP to produce inorganic phosphorus. The activity of ATPase can be expressed by measuring the production amount of inorganic phosphorus in unit time. In the control system, Ca2+ -ATPase activity was inhibited, while in the sample system, Ca2+ -ATPase activity was not inhibited. The difference of inorganic phosphorus content between the sample and the control was the inorganic phosphorus produced by Ca2+ -ATPase during the incubation time. The activity of Ca2+ -ATPase was determined by inorganic phosphorus production.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 10 mL x 1 vial | 20 mL x 1 vial | 2-8°C, 12 months |
| Reagent 2 | Activator A | 1 mL x 1 vial | 2 mL x 1 vial | 2-8°C, 12 months |
| Reagent 3 | Activator B | 1 mL x 1 vial | 2 mL x 1 vial | 2-8°C, 12 months |
| Reagent 4 | Substrate | Powder x 1 vial | Powder x 1 vial | 2-8°C, 12 months |
| Reagent 5 | Protein Precipitator | 3 mL x 1 vial | 6 mL x 1 vial | 2-8°C, 12 months |
| Reagent 6 | Chromogenic Agent A | Powder x 1 vial | Powder x 2 vials | 2-8°C, 12 months shading light |
| Reagent 7 | Acid Reagent | 5 mL x 1 vial | 10 mL x 1 vial | 2-8°C, 12 months |
| Reagent 8 | Chromogenic Agent B | Powder x 1 vial | Powder x 2 vials | 2-8°C, 12 months |
| Reagent 9 | 10 mmol/L Standard Solution | 2 mL x 1 vial | 2 mL x 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 rat liver 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/kg wet weight) | 5.60 | 85.40 | 195.00 |
| %CV | 6.5 | 6.1 | 5.4 |
Inter-assay Precision
Three rat liver samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (U/kg wet weight) | 5.60 | 85.40 | 195.00 |
| %CV | 7.6 | 8.2 | 8.2 |
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 107%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/kg wet weight) | 25.8 | 103 | 242 |
| Observed Conc. (U/kg wet weight) | 28.4 | 111.2 | 249.3 |
| Recovery rate (%) | 110 | 108 | 103 |
Sensitivity
The analytical sensitivity of the assay is 1.18 U/kg wet weight. 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.