PKM2 ELISA Kit (Pyruvate Kinase Isozymes M1/M2)

INTENDED USE

Human PKM2 ELISA kit can measure concentrations of pyruvate kinase isozymes M1/M2 (PKM2, p58, PK2, TCB, THBP1, tumor M2-PK, Thyroid hormone-binding protein 1) present is plasma, serum, biological fluids or tissue homogenate samples. This assay has a sensitivity detection limit of 46.875 pg/ml.

INTRODUCTION

PKM2, which stands for Pyruvate kinase isozymes M1/M2 is an enzyme, in a metabolic process called glycolysis. Glycolysis breaks down glucose to produce energy in the form of ATP. There are two forms of pyruvate kinase; M1 and M2. The M1 form is found in adult tissues. Is always active. On the hand PKM2 is found in tissues and certain cancer cells.

Unlike PKM1 PKM2 has the ability to switch between an dimeric form and a more active tetrameric form. This allows it to control the flow of glycolysis. The tetramerization of PKM2 depends on signals from oncogenes and nutrient availability. When there are nutrients PKM2 forms tetramers, leading to glycolytic activity and ATP production. However in cancer cells oncogenic signaling pathways cause PKM2 to dimerize instead slowing down glycolysis and redirecting metabolic intermediates towards producing nucleotides, amino acids and lipids that support cell growth.

In addition to its role in regulating glycolysis dimeric PKM2 can move into the nucleus. Function as a protein kinase, by adding groups to various proteins involved in gene expression regulation, cell proliferation and even glycolysis itself. Nuclear PKM2 also interacts with hypoxia factor 1 (HIF 1) to activate genes related to glucose metabolism and angiogenesis.

Furthermore the dimeric form of PKM2 shows a decrease, in its function. This leads to the accumulation of intermediates upstream, which are then redirected towards the pentose phosphate and serine biosynthesis pathways. The purpose of this redirection is to generate cofactors and amino acids meeting the antioxidant needs of rapidly dividing tumor cells. As a result PKM2 plays a role, in reshaping the metabolic processes of cancer cells.

CONTENT

All reagents supplied need to be stored at 2 °C – 8 °C, unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date.

• One 96-Well Plate: Pre-coated with anti- PKM2.
• Standards: Lyophilized recombinant.
• Sample/Standard Dilution Buffer.
• Biotinylated-labelled Antibody.
• Antibody Dilution Buffer.
• HRP-Streptavidin Conjugate (SABC).
• SABC Dilution Buffer.
• TMB Substrate.
• Wash Buffer (25x).
• Plate Sealer.
• Product Instructions.

TYPICAL RESULTS

For this PKM2 ELISA kit, it is recommended that a standard curve is generated for each assay carried out.

Standard Curve: 0, 78.125, 156.25, 312.5, 625, 1250, 2500, 5000 pg/ml.
Reactivity: Human
Sensitivity: 46.875 pg/ml
Range: 78.125 – 5000 pg/ml
Principle: Sandwich, Double Antibody
Application: Research Use Only.

ASSAY CHARACTERISTICS

– Specificity: Highly specific for PKM2, no cross reactivity or interference between PKM2 and analogues was detected.
– Recovery: Serum (88 – 103%), EDTA Plasma (91 – 102%), Heparin Plasma (91 – 100%).
– Linearity: Serum (93 – 104%), EDTA Plasma (89 – 99%), Heparin Plasma (83 – 99%).
– Precison Intra-Assay: CV < 8%
– Precison Inter-Assay: CV < 10%

REFERENCES

1. Pyruvate kinase M2 (PKM2) in cancer and cancer therapeutics. Cancer Lett. 2021 Apr 10;503:240-248. Zhu S., et al.
2. Pyruvate kinase deficiency. Clin Biochem. (1990) 23 (2): 155-7. Miwa S. and Fujii H.
3. Myeloid Cell PKM2 Deletion Enhances Efferocytosis and Reduces Atherosclerosis. Circ Res. (2022) 130 (9): 1289-1305. Doddapattar P., et al.

ADDITIONAL INFORMATION

• Full Name: PKM2 ELISA Kit (Pyruvate Kinase Isozymes M1/M2)
• Reactivity: Human
• Sample Type: Plasma, Tissue Homogenates, Serum, Biological Fluids
• Sensitivity: 46.875 pg/ml

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