ELISA Kit for Pyruvate kinase isozymes M2 (PKM2)

M2PK; PKM; M2-PK; PKM1; CTHBP; OIP3; PK3; PKM; TCB; THBP1; Pyruvate kinase muscle isozyme; Thyroid hormone-binding protein 1; Cytosolic thyroid hormone-binding protein; Pyruvate Kinase, Muscle

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.

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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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