Description
Key facts
Detection method:Fluorescent,
Sample types:Cell culture extracts, Tissue Extracts, Cell Lysate,
Assay type:Quantitative,
Sensitivity:= 40 nM,
Assay time:1h,
Assay Platform:Microplate reader
Product details:
Coenzyme A Assay Kit (Fluorometric - Green) (ab138889) provides an ultrasensitive fluorometric assay to quantitate CoA content by detection of the -SH group in CoA.
The fluorogenic CoA green indicator dye used in the kit becomes strongly fluorescent upon reacting with -SH.
The assay kit can detect as little as 4 pmol CoA/ 100 uL assay volume (40 nM). It can be performed in a convenient 96-well or 384-well microtiter-plate format at Ex/Em = 490/520 nm, and easily adapted to automation without a separation step.
Coenzyme A assay protocol summary:
- add samples and standards to wells
- add reaction mix
- incubate for 10-60 min whilst measuring fluorescence with a microplate reader
The kit is for solution-based inhibitors or activators of Coenzyme A only. Any samples that contain proteins or thiol-containing compounds are not compatible with the kit.
Other Notes
Previously called Coenzyme A Detection kit (Fluorometric - Green).
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.
Coenzyme A (CoA) is an important enzyme cofactor with a molecular mass of approximately 767.5 Da. Alternate names include CoA-SH and acetyl coenzyme A when bonded to an acetyl group. CoA is widely expressed in both prokaryotic and eukaryotic cells including the liver heart and brain tissues playing a significant role in metabolic processes. It functions mechanically by carrying acyl groups such as acetate which is vital for the conversion of pyruvate in the Krebs cycle and fatty acid synthesis.
Biological function summary
This cofactor is important for various enzymatic reactions especially those involving the metabolism of carbohydrates fats and amino acids. It is not a part of a larger protein complex but works closely with enzymes like acetyl-CoA carboxylase. Its ability to form energy-rich thioester bonds allows for the transfer of acyl groups making it essential for energy production and biosynthetic reactions in cells. CoA transfers these acetyl groups in forms like acetyl coenzyme A or acetyl CoA which are critical for cellular functions.
Pathways
CoA is essential in both the citric acid cycle (Krebs cycle) and fatty acid metabolism. In the citric acid cycle it helps convert pyruvate into acetyl-CoA therefore linking glycolysis to energy production. In fatty acid metabolism CoA participates in the process of beta-oxidation. Both pathways involve enzymes such as pyruvate dehydrogenase and fatty acid synthase which collaborate with CoA in important steps.
Perturbations in CoA metabolism are associated with conditions such as neurodegenerative diseases and metabolic disorders like diabetes. Alterations in CoA levels may affect pathways involving proteins such as pyruvate dehydrogenase and acetyl-CoA carboxylase which are important in maintaining cellular energy balance and lipid metabolism. These links highlight the importance of CoA in health and disease contexts.
