ELISA Kit for Pyruvate kinase isozymes M2 (PKM2)
Cat# SEA588Mu-96x10
Size : 96Tx10
Brand : Cloud-Clone Corp
Specificity
This assay has high sensitivity and excellent specificity for detection of Pyruvate kinase isozymes M2 (PKM2).
No significant cross-reactivity or interference between Pyruvate kinase isozymes M2 (PKM2) and analogues was observed.
Recovery
Matrices listed below were spiked with certain level of recombinant Pyruvate kinase isozymes M2 (PKM2) and the recovery rates were calculated by comparing the measured value to the expected amount of Pyruvate kinase isozymes M2 (PKM2) in samples.
| Matrix | Recovery range (%) | Average(%) |
| serum(n=5) | 95-103 | 99 |
| EDTA plasma(n=5) | 79-104 | 91 |
| heparin plasma(n=5) | 82-103 | 86 |
Precision
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Pyruvate kinase isozymes M2 (PKM2) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Pyruvate kinase isozymes M2 (PKM2) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Pyruvate kinase isozymes M2 (PKM2) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
| Sample | 1:2 | 1:4 | 1:8 | 1:16 |
| serum(n=5) | 89-96% | 91-105% | 91-99% | 78-102% |
| EDTA plasma(n=5) | 94-103% | 83-101% | 98-105% | 89-102% |
| heparin plasma(n=5) | 82-91% | 79-102% | 94-105% | 97-105% |
Stability
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
Reagents and materials provided
| Reagents | Quantity | Reagents | Quantity |
| Pre-coated, ready to use 96-well strip plate | 1 | Plate sealer for 96 wells | 4 |
| Standard | 2 | Standard Diluent | 1×20mL |
| Detection Reagent A | 1×120µL | Assay Diluent A | 1×12mL |
| Detection Reagent B | 1×120µL | Assay Diluent B | 1×12mL |
| TMB Substrate | 1×9mL | Stop Solution | 1×6mL |
| Wash Buffer (30 × concentrate) | 1×20mL | Instruction manual | 1 |
Assay procedure summary
1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.