Description
Key facts
Detection method:Colorimetric,
Sample types:Tissue Lysate, Cell Lysate,
Assay type:Enzyme activity,
Sensitivity:< 1 mU/well,
Range:2 - 10 nmol/well,
Assay time:1h 30m,
Assay Platform:Microplate reader
Product details:
Phosphofructokinase Activity Assay kit ab155898 is a mix-and-read assay to quantify 6-PFK activity. Readout on any colorimetric (450 nm) plate reader.
PFK activity assay principle
In the PFK activity assay protocol, PFK (phosphofructokinase) converts fructose-6-phosphate and ATP to fructose-diphosphate and ADP. The ADP in the presence of substrate and enzyme mix is converted to AMP and NADH, which reduces a colorless probe to a colored product with strong absorbance at 450 nm.
PFK activity assay protocol summary
- add samples and standards to wells
- add reaction mix
- analyze with a microplate reader over 20-60 min
Other notes
This product was previously called K776 Biovision Phosphofructokinase (PFK) Activity Colorimetric Assay Kit and 6-Phosphofructokinase Activity Assay Kit (Colorimetric). Biovision was acquires by Abcam in 2021.
Phosphofructokinase (PFK) is a key glycolytic enzyme and plays a major regulatory role during glycolysis. This enzyme is present in bacteria, plants and animals. There are 2 types of PFKs - PFK1 and PFK2. In the presence of ATP, PFK1 & PFK2 catalyzes the conversion of fructose-6-phosphate to fructose-1,6-diphosphate and fructose-2,6-diphosphate respectively and ADP. PFK has 3 major isoforms in mammals: PFK-M (muscle), PFK-L (liver) and PFK-P (platelet).
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
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.
6-Phosphofructokinase often abbreviated as PFK or PFK-1 is an important enzyme in the glycolytic pathway catalyzing the phosphorylation of fructose-6-phosphate to fructose-16-bisphosphate. This reaction is a major rate-limiting step in glycolysis controlling the pace of glucose breakdown for energy production. PFK-1 is a tetrameric enzyme with a molecular mass of about 340 kDa. It is expressed in a variety of tissues including muscle liver and brain reflecting its essential role in energy metabolism across different tissues.
Biological function summary
This enzyme plays an important role in regulating glycolysis where it ensures efficient energy production under varying cell conditions. PFK-1 acts as a metabolic sensor responding to cellular energy demands by allosteric regulation through ATP ADP AMP and citrate. By forming part of a multisubunit complex it regulates substrate flux within glycolysis and changes in PFK-1 activity directly affect how cells respond to energy needs and stress.
Pathways
6-phosphofructokinase functions critically within glycolysis and by extension influences the TCA cycle. The activity of this enzyme links to key metabolites like ATP and AMP which regulate its function. Furthermore PFK-1 activity relates to the activity of enzymes such as hexokinase and pyruvate kinase forming a coordinated system that controls the rate of glucose oxidation and energy yield highlighting its pivotal role in cellular metabolic networks.
PFK-1 has connections to conditions such as cancer and glycogen storage disease type VII also known as Tarui's disease. In cancer enhanced glycolytic flux often accompanies increased PFK-1 activity supporting tumour growth. In Tarui's disease mutations in the PFKM gene lead to deficient PFK-1 activity in muscle causing impaired glycogen metabolism. This enzyme's dysregulation can lead to significant metabolic disturbances making it a target for therapeutic interventions in such conditions.
