Pyruvate kinase PKM Antibodies

Pyruvate kinase PKM (PKM)

Glycolytic enzyme that catalyzes the transfer of a phosphoryl group from phosphoenolpyruvate (PEP) to ADP, generating ATP. Stimulates POU5F1-mediated transcriptional activation.

Plays a general role in caspase independent cell death of tumor cells. The ratio between the highly active tetrameric form and nearly inactive dimeric form determines whether glucose carbons are channeled to biosynthetic processes or used for glycolytic ATP production.

Gene Information

Human
Gene Name:	PKM
Uniprot:	P14618
Entrez:	5315

Family

Belongs to:
pyruvate kinase family

Alternative Names

CTHBP; Cytosolic thyroid hormone-binding protein; MGC3932; OIP3; OIP-3; OIP3EC 2.7.1.40; Opa-interacting protein 3; p58; PK, muscle type; PK2; PK3; PK3PKM; PKM2; Pyruvate kinase 2/3; pyruvate kinase isozymes M1/M2; Pyruvate kinase muscle isozyme; pyruvate kinase, muscle; TCB; THBP1; THBP1p58; Thyroid hormone-binding protein 1; thyroid hormone-binding protein, cytosolic; Tumor M2-PK

Molecular Weight

Mass (kDA):
57.937 kDA

Genomic Context

Human
Location:	15q23
Sequence:	15; NC_000015.10 (72199029..72231624, complement)

Tissue Specificity

Specifically expressed in proliferating cells, such as embryonic stem cells, embryonic carcinoma cells, as well as cancer cells.

Subcellular Localizations

Cytoplasm. Nucleus. Translocates to the nucleus in response to different apoptotic stimuli. Nuclear translocation is sufficient to induce cell death that is caspase independent, isoform-specific and independent of its enzymatic activity.

Diseases

• Malignant Neoplasms
• Neoplasms
• Acute Promyelocytic Leukemia
• Liver Carcinoma
• Salmonella Infections
• Cell Transformation, Neoplastic
• Hemorrhage
• Neuroblastoma
• Glioma
• Ischemia
• Depressive Disorder
• Hypoxia

• Sarcoma
• Mental Disorders
• Tissue Adhesions
• Systemic Infection
• Migraine Disorders
• Salmonella Infections, Animal
• Sclerosis

Interacting Proteins

• ARAF
• ARRB1
• ARRB2
• CTNNB1
• DAPK1

• EGLN3
• HIF1A
• HIST1H3A
• HTT
• MAPK3

• RAF1
• TINF2

Pathways

• Proteolysis
• Glycolysis
• Long-term Memory
• Localization
• Oxidative Phosphorylation
• Synaptic Transmission
• Cell Differentiation
• Oncogenesis
• Cell Proliferation
• Autolysis
• Pathogenesis
• Transport
• Protein Phosphorylation

• Fertilization
• Brain Development
• Muscle Contraction
• Sensitization
• Translation
• Phagocytosis
• Cell Cycle

PTMs

• Phosphorylation
• Cleavage

• Myristoylation
• Dephosphorylation

• Acetylation

Research Areas

• Cancer
• Lipid and Metabolism

For details, visit:
bosterbio.com/bosterbio-gene-info-cards/PKM

Boster Bio - Best Uses Of The PKM Marker

This article will explore the most effective uses for the Boster Bio PKM marker. We will also discuss Electrophoresis as well as Chemiluminescence. After we've covered these two methods, we'll discuss the Boster Bio PKM Marker in general. Here are other methods we use in our work. They can be helpful if you're looking to identify the exact protein or gene in a sample.

Electrophoresis

The protein is produced by cells throughout a variety of organs. PKM levels that are high PKM are associated with a poor prognosis for several cancers, including liver hepatocellular tumor, pancreatic adenocarcinoma, head and neck squamous cell carcinoma, and thyroid cancer. A high level of PKM are associated positively with prognoses for several other cancers, including stomach, lung, and pancreatic.

PKM2 interacts with Oct-4, a nuclear protein that is involved in self-renewal as well as differentiation of cancer cells. HIF-1, a coactivator for transcriptional activation in the nucleus, facilitates this interaction. PKM2-expressing cells are more likely have cancer-related genes. This suggests that PKM2 could be an effective therapeutic target or diagnostic marker for the disease.

In addition to phosphorylated SNAP-23, PKM2 also binds to PKM2. This protein may be associated with SNAP23 during docking at the plasma membrane. The marker PKM is expressed in a wide variety of cell types and the results of these studies suggest this protein may promote exocytosis in cells. This relationship is not fully understood.

PKM2 is an enzyme that plays an essential role in the glycolysis metabolism during exocytosis of tumor cells. There are four different forms of the molecule. The first, PKM1, is expressed during pregnancy. The second, PKM2, is expressed in adulthood. The expression of PKM2 increases the amount of glucose and reduces the consumption of O2 in cancer cells.

PKM2 is also available in dimer form. There are two forms of PKM2 that are highly active, the Tetramer as well as the low-activity Dimer. Both are controlled by multiple molecules, like E7 oncoproteins and Prylrylation-mediated Tyrosine Kinase.

Recombinant PKM2 was incubated with SNAP-23 in a kinase buffer which contains 50 mM Tris-HCl (pH 7.5), 100 mg KCl, 1 mM Na3VO4, and 0.5 mg PEP. The recombinant protein was loaded onto SDS–PAGE.

Proteins were separated using SDS-PAGE and stained. The particles were then resuspended 0.1% formic acid, and incubated at 56 degrees C for 30 minutes. The samples were then subjected to a 56-degree temperature incubation for 30 minutes. After that 200 millimeters IAA was added to the mixture , and then left in the dark for 20 mins. This was to aid in alkylation. The gels were then examined with liquid chromatography, and mass spectrometry/MS/MS.

PKM2 also plays a part in the release of exosomes out of cancer cells. It phosphorylates SNAP-23 to enable exosome secretion. This process is strongly associated with the metabolic switch between oxidative phosphorylation and aerobic glycolysis. It may be caused by a perturbation of PKM2. Further, PKM2 expression is closely linked with the increased release of exosomes.

Chemiluminescence

For studies of immunohistochemistry, Boster Bio's ECL Plus Western Blotting Substrate is a luminol-based and ultra-sensitive chemiluminescent substance that detects horseradish peroxidase. It can be applied to nitrocellulose and PVDF membranes and is compatible with a variety of blocking buffers. The reagent produces low background levels and is suitable for imaging on x-ray film as well as CCD imaging systems. For a more accurate visualization this substrate can be used in conjunction with a shaking platform during the process of incubation.

HUVECs were developed on glass coverslips six well plates containing 0.1% Triton X100. The cells were then kept at 37°C for 2 hours in 5 percent CO2. The cells were washed twice using Hank's solution, to get rid of nonadherent cells and measured using a Bio-Rad Model 550.

ELISA Multiplex kits can be loaded with samples of 50 ul per well, and 18 targets can be quantified. Multiplex ELISA kits contain highly specific spots for antibodies that are printed in the appropriate locations on each well's surface. Boster Biological Technology's expert team is responsible for ensuring the accuracy of the printing and separates specific spots to increase the sensitivity of ELISA. ELISA Multiplex ELISA kits are available in a variety of concentrations. They also have high sensitivity and a Lower Limit of Detection.

The paper-based devices were released over a decade ago. Their unique combination of features made them ideal for applications on-site. This technology is suitable for point-of-care diagnostics due to its paper-based format and the chemiluminescent technology. They are not widely utilized at the moment. In the near future, they could be the standard for testing cancer markers. For the time being it is unlikely that it will replace the conventional colorimetric methods.