Creatine Kinase (CK) Activity Assay Kit
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Creatine Kinase (CK) Activity Assay Kit
| SKU # | E-BC-K558-S |
| Detection Instrument | Spectrophotometer (340 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | CK, Creatine phosphokinase |
| Sample type | Serum, plasma, animal tissue, cells |
| Sensitivity | 3.7 U/L |
| Detection range | 3.7-160 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 80 min |
| Precision | Average inter-assay CV: 8.100% | Average intra-assay CV: 5.200% |
| Other instruments required | Micropipettor, Incubator, Vortex mixer |
| Other reagents required | PBS (0.01 M, pH 7.4) |
| 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  (3.7-160 U/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Human plasma | 1 |
| Mouse serum | 1 |
| Rat serum | 1 |
| HepG2 cells homogenization | 1 |
| 10% Rat kidney tissue homogenization | 1 |
| 10% Rat brain tissue homogenization | 2-5 |
| 10% Rat liver tissue homogenization | 2-10 |
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Note: The diluent is  PBS (0.01 M, pH 7.4).
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Detection Principle
Creatine kinase (CK) catalyze creatine phosphate and ADP to produce creatine and ATP. Hexokinase catalyze creatine and glucose to produce glucose-6-phosphate. Glucose-6-phosphate dehydrogenase (G-6-PD) catalyze glucose-6-phosphate and NADP+ to produce NADPH which have a maximum absorption peak at 340 nm. The CK activity can be calculated by measuring the OD values at 340 nm.
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Kit Components & Storage
| Item | Component |
Size 1 (50 assays) |
Size 2 (100 assays) |
Storage |
| Reagent 1 | Enzyme Solution | 60 mL × 1 vial | 60 mL × 2 vials | 2-8℃, 12 months shading light |
| Reagent 2 | Acid Solution | 6 mL × 1 vial | 12 mL × 1 vial | 2-8℃, 12 months shading light |
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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.
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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/L) | 8.4 | 26.8 | 95.5 |
| %CV | 5.6 | 4.9 | 5.1 |
Inter-assay Precision
Three human serum samples were assayed 17 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μmol/L) | 8.4 | 26.8 | 95.5 |
| %CV | 8.4 | 7.9 | 8.0 |
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Sensitivity
The analytical sensitivity of the assay is 3.7 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.
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Standard Curve
Take rat muscle as an example, take 2% rat muscle homogenate 50μL, operate according to the operation steps, record the OD value A1, react at 37℃, measure the measuring tube every minute, record the OD value:
| Reaction time (min) | 0 | 1 | 2 | 3 | 4 | 5 | 6 |
| Absoluted OD | 0.427 | 0.451 | 0.473 | 0.496 | 0.520 | 0.541 | 0.566 |