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). In humans, PFK deficiency causes glycogen storage disease, also called Tarui's disease, which is characterized by exercise-induced muscle weakness and cramps. On the other hand, increased PFK activity contributes to cancer cell proliferation and tumorigenicity. Early detection of abnormal phosphofructokinase activity is crucial for diagnosis, prediction and therapeutic strategy. In Assay Genie's Phosphofructokinase Activity Assay kit, PFK 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 is simple, robust, and rapid and can detect phosphofructokinase activity less than 1 mU.

Figure 1: (a).NADH standard curve (b). Phosphofructokinase activity in bovine liver and HeLa cell lysate. Assays were performed following kit protocol.