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ELISA Kit for Pyruvate kinase isozymes M2 (PKM2)
ELISA Kit for Pyruvate kinase isozymes M2 (PKM2)
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Enzyme-linked immunosorbent assay for Antigen Detection.
Product No.
SEA588Hu
Organism Species
Homo sapiens (Human).
Sample Type
Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Test Method
Double-antibody Sandwich
Assay Length
3h
Detection Range
78-5,000pg/mL
Sensitivity
The minimum detectable dose of this kit is typically less than 29pg/mL.
UOM
48T 96T 96T*5 96T*10 96T*100
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) | 80-89 | 85 |
| EDTA plasma(n=5) | 81-96 | 90 |
| heparin plasma(n=5) | 91-105 | 95 |
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) | 88-96% | 78-105% | 79-97% | 87-101% |
| EDTA plasma(n=5) | 95-105% | 99-105% | 90-103% | 92-101% |
| heparin plasma(n=5) | 88-97% | 81-99% | 94-103% | 93-102% |
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.
| Magazine | Citations |
| Cancer Research | Antigen Presentation by Dendritic Cells in Tumors Is Disrupted by Altered Metabolism that Involves Pyruvate Kinase M2 and Its Interaction with SOCS3 AACR: 70189 |
| Molecular BioSystems | Abnormal levels of heterogeneous nuclear ribonucleoprotein A2B1 (hnRNPA2B1) in tumour tissue and blood samples from patients diagnosed with lung cancer Pubmed:25483567 |
| Tumour Biol | M2 isoform of pyruvate kinase (PKM2) is upregulated in Kazakh’s ESCC and promotes proliferation and migration of ESCC cells PubMed: 26404132 |
| Oncotarget | Identification of a serum biomarker panel for the differential diagnosis of cholangiocarcinoma and primary sclerosing cholangitis Pubmed:29707118 |
| Journal of Neurotrauma | Acteoside improves muscle atrophy and motor function by inducing new myokine secretion in chronic spinal cord injury Pubmed: 30318996 |
| Cell Reports | Secreted Pyruvate Kinase M2 Promotes Lung Cancer Metastasis through Activating the Integrin Beta1/FAK Signaling Pathway Pubmed: 32049010 |
| Chrysin serves as a novel inhibitor of DGKα/FAK interaction to suppress the malignancy of esophageal squamous cell carcinoma (ESCC) | |
| Cancer Cell International | Bufalin Induced Mitochondrial Dysfunction Promotes Apoptosis of Glioma Cells by Regulating Annexin A2 and DRP1 Proteins |
| Cancer Cell Int | Bufalin induces mitochondrial dysfunction and promotes apoptosis of glioma cells by regulating Annexin A2 and DRP1 protein expression 34376212 |
| Life (Basel) | Putative Association between Low Baseline Gene Expression in the Peripheral Blood and Clinical Remission in Rheumatoid Arthritis Patients Treated with Tofacitinib 34947916 |
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