Abcam, ab83432, Pyruvate Kinase Assay Kit

Size: 100Test / 2000Test
Pyruvate Kinase Assay Kit ab83432 is a no-wash assay with one 15 minute incubation at 25°C. In the assay, pyruvate kinase acts on PEP and ADP to generate pyruvate, which is then oxidized. Readout is on any colorimetric (570 nm) or fluorometric (Ex/Em 535/587 nm) plate reader. - Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs. A Pyruvate Kinase Activity Assay Kit is used in the study and diagnosis of diseases where glycolytic dysfunction is involved, such as pyruvate kinase deficiency, a hereditary cause of hemolytic anemia, and various cancers, where altered PK activity supports tumor metabolism. It's also relevant in metabolic and liver disorders, neonatal anemia, and toxicological studies, helping assess cellular energy production and metabolic disruption.
Key facts
Detection method:Colorimetric/Fluorometric,
Sample types:Plasma, Tissue, Cell culture supernatant, Serum, Other biological fluids, Adherent cells, Suspension cells,
Assay type:Enzyme activity (quantitative),
Sensitivity:= 0.1 mU/mL,
Assay time:40m,
Assay Platform:Microplate reader

Product details:
Pyruvate Kinase Assay Kit ab83432 provides a simple, direct and automation-ready assay for measuring pyruvate kinase activity in cell culture supernatant, plasma, serum, tissue, urine and other biological fluids (not validated for use with whole blood).
How the assay works
In the pyruvate kinase assay protocol, PEP and ADP are catalyzed by pyruvate kinase to generate pyruvate and ATP. The generated pyruvate is oxidized by pyruvate oxidase to produce color (absorbance 570 nm) and fluorescence (Ex/Em 535/587 nm). Since the increase in color or fluorescence intensity is proportional to the increase in pyruvate amount, the PK activity can be accurately measured.
The kit detects 0.1 mU/ml pyruvate kinase.
Pyruvate kinase assay protocol summary
- Add samples and standards to wells
- Add reaction mix
- Analyze with microplate reader immediately and after 10-20 min
Other notes
This product is manufactured by BioVision, an Abcam company and was previously called K709 Pyruvate Kinase Activity Colorimetric/Fluorometric Assay Kit. K709-100 is the same size as the 100 test size of ab83432.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions--20°C, Appropriate long-term storage conditions--20°C, Storage information--20°C

Supplementary Information:
This supplementary information is collated from multiple sources and compiled automatically.
Pyruvate kinase (PK) also known as ATP: pyruvate 2-O-phosphotransferase is an enzyme fundamental to the glycolytic pathway. This enzyme catalyzes the final step where phosphoenolpyruvate (PEP) is converted to pyruvate yielding one molecule of ATP. Pyruvate kinase has several isoforms including PKM1 and PKM2 with molecular weights ranging around 57-58 kDa. These isoforms are expressed in various tissues; for example PKM1 is found in muscle tissue while PKM2 occurs in proliferating tissues and certain tumors.
Biological function summary
Pyruvate kinase plays an important role in cellular metabolism by regulating the rate of glycolysis. It is not part of a complex but functions independently to facilitate the conversion of PEP to pyruvate. This conversion is important for energy production particularly in tissues where ATP is in high demand. Pyruvate kinase activity is regulated by a variety of metabolites enhancing its function under different physiological conditions.
Pathways
Pyruvate kinase serves an integral part of the glycolytic pathway. It links to key metabolic pathways such as gluconeogenesis and is influenced by proteins like lactate dehydrogenase and fructose-16-bisphosphate which serve to further manage its activity. The enzyme is a point of regulation within the glycolytic pathway affecting the overall energy management within cells especially under varying oxygen levels.
Altered pyruvate kinase activity associates with conditions like pyruvate kinase deficiency and cancer. Pyruvate kinase deficiency often related to mutations impacting PKLR or PKM genes disrupts ATP production in red blood cells leading to hemolytic anemia. In various cancers especially where PKM2 is upregulated the enzyme's altered function contributes to the metabolic reprogramming of cancer cells. In these diseases PKM2 often interacts with other proteins such as hypoxia-inducible factor 1-alpha (HIF-1α) playing a role in adapting to hypoxic conditions common in tumors.