Pyruvate Kinase (PK) Activity Assay Kit
Pyruvate Kinase (PK) Activity Assay Kit
| SKU # | E-BC-K611-M |
| Detection Instrument | Microplate reader (330-350 nm) |
Product Details
Properties
| Synonyms | PK |
| Sample type | Serum, plasma, tissue and cell |
| Sensitivity | 0.08 U/L |
| Detection range | 0.08–6.79 U/L |
| Assay type | Enzyme Activity |
| Assay time | 10 min |
| Precision | Average inter-assay CV: 7% | Average intra-assay CV: 5% |
| Other instruments required | Test tube, Micropipettor, Vortex mixer, Incubator |
| Storage | -20℃ |
| 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 kidney tissue homogenate | 4-6 |
| 10% Rat lung tissue homogenate | 4-6 |
| 10% Rat liver tissue homogenate | 4-6 |
| 10% Rat brain tissue homogenate | 4-6 |
| 10% Mouse liver tissue homogenate | 4-6 |
| 10% Mouse heart tissue homogenate | 4-6 |
| Rat serum | 1 |
| Rat plasma | 1 |
| Mouse serum | 1 |
| Mouse plasma | 1 |
| Human serum | 1 |
| Dog plasma | 1 |
| HL-60 cell | 1 |
| 293T cell | 1 |
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
In the presence of adenosine diphosphate (ADP), pyruvate kinase (PK) catalyzes the conversion of phosphoenolpyruvate (PEP) to pyruvate, Lactate dehydrogenase catalyzes the reaction of pyruvic acid with NADH to produce lactic acid and NAD+. The activity of pyruvate kinase can be calculated by detection the absorbance changes at 340 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 10 mL ×1 vial | 10 mL × 2 vials | -20°C, 12 months |
| Reagent 2 | Substrate A | Powder × 1 vial | Powder × 2 vials | -20°C, 12 months shading light |
| Reagent 3 | Substrate B | Powder × 1 vial | Powder × 2 vials | -20°C, 12 months shading light |
| Reagent 4 | Enzyme Solution | 1.2 mL × 1 vial | 1.2 mL × 2 vials | -20°C, 12 months shading light |
| UV 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 (U/L) | 1.20 | 3.40 | 5.80 |
| %CV | 5.35 | 5.0 | 4.7 |
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 (U/L) | 1.20 | 3.40 | 5.80 |
| %CV | 6.8 | 7.3 | 6.9 |
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 103%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/L) | 1.8 | 3.7 | 6.2 |
| Observed Conc. (Ul/L) | 1.8 | 3.9 | 6.5 |
| Recovery rate (%) | 99 | 105 | 105 |
Sensitivity
The analytical sensitivity of the assay is 0.08 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.