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 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 450nm. PFK activity assay is simple, robust, and rapid and can detect phosphofructokinase activity less than 1mU.
Sample Type:
Animal tissues, cell cultures.
Intended Use:
Measurement of phosphofructokinase activity in various tissues/cells. Analysis of glucose metabolism and cell signaling in various cell types. Screening anti-cancer drugs.
Sensitivity:
can detect PFK activity at <1mU.
Kit Components:
PFK assay buffer, 1x27ml
PFK substrate (lyophilized), 1 vial
ATP (lyophilized), 1 vial
PFK enzyme mix (lyophilized), 1 vial
PFK developer (lyophilized), 1 vial
NADH standard (lyophilized), 1 vial
Positive control (lyophilized), 1 vial
Storage and Stability:
Store powder at 4 degrees C liquid at -20 degrees C. Store other components at 4 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.