Skip to main content

Phosphofructokinase Activity Assay Kit (6-PFK, Colorimetric) ab155898 is simple, robust, and rapid and can detect phosphofructokinase activity less than 1 mU.

Be the first to review this product! Submit a review

Images

Functional Studies - Phosphofructokinase Activity Assay Kit (6-PFK, Colorimetric) (AB155898), expandable thumbnail

Publications

Key facts

Detection method
Colorimetric
Sample types
Tissue Lysate, Cell Lysate
Assay type
Enzyme activity
Reactive species
Mammals
Range
2 - 10 nmol/well
Assay time
1h 30m
Sensitivity
< 1 mU/well

Target data

Function

Catalyzes the phosphorylation of D-fructose 6-phosphate to fructose 1,6-bisphosphate by ATP, the first committing step of glycolysis (PubMed:22923583). Negatively regulates the phagocyte oxidative burst in response to bacterial infection by controlling cellular NADPH biosynthesis and NADPH oxidase-derived reactive oxygen species. Upon macrophage activation, drives the metabolic switch toward glycolysis, thus preventing glucose turnover that produces NADPH via pentose phosphate pathway (By similarity).

Additional Targets

PFKM, PFKP

Alternative names

What's included?

100 Test
Components
ATP III
1 x 1 Vial
Assay Buffer 7
1 x 25 mL
CK Enzyme Mix
1 x 1 Vial
Developer Solution III
1 x 1 Vial
NADH Standard I
1 x 1 Vial
PFK Positive Control
1 x 1 Vial
PFK Substrate
1 x 1 Vial

Recommended products

Phosphofructokinase Activity Assay Kit (6-PFK, Colorimetric) ab155898 is simple, robust, and rapid and can detect phosphofructokinase activity less than 1 mU.

Key facts

Detection method
Colorimetric
Sample types
Tissue Lysate, Cell Lysate
Assay type
Enzyme activity
Reactive species
Mammals
Range
2 - 10 nmol/well
Assay time
1h 30m
Assay Platform
Microplate reader
Sensitivity
< 1 mU/well

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C

Notes

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

1 product image

  • Functional Studies - Phosphofructokinase Activity Assay Kit (6-PFK, Colorimetric) (ab155898), expandable thumbnail

    Functional Studies - Phosphofructokinase Activity Assay Kit (6-PFK, Colorimetric) (ab155898)

    PFK activity determined in bovine liver and HeLa cell lysate. Assay performed following kit protocol.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com