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Creatine Kinase (CK) Activity Assay Kit
SKU: E-BC-K558-S-100
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 |
Note: The diluent is PBS (0.01 M, pH 7.4).
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.
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 |
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/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 |
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.
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 |