Description
Key facts
Detection method:Colorimetric,
Sample types:Tissue, Cell Lysate,
Assay Platform:Microplate reader
Product details:
Citrate Synthase Assay Kit (ab239712) provides reagents for the measurement of citrate synthase activity in various tissues/cells. The assay can detect Citrate Synthase activity less than 1 mU in a variety of samples.
Citrate synthase assay principle
In the citrate synthase assay, citrate synthase acts on acetyl-CoA and oxaloacetate to produce citrate + CoA-SH. CoA-SH then reacts with DTNB to produce a colorimetric (412 nm) readout.
Citrate synthase assay protocol summary
- add samples and standards to wells
- add reaction mix
- Analyse on a colorimetric plate reader immediately in kinetic mode at room temperature for 20-40 mins.
Related and recommended products
This kit uses the same method as Citrate synthase activity assay kit
ab119692
, except without the antibody pull-down step used in
ab119692
to isolate citrate synthase.
Other Notes
This product was previously called K318 Biovision Citrate Synthase Activity Colorimetric Assay Kit. Biovision was acquired by Abcam in 2021.
REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
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.
Citrate synthetase also known as citrate synthase is an important enzyme in the tricarboxylic acid cycle. It catalyzes the condensation of acetyl-CoA and oxaloacetate to form citrate and CoA. The enzyme weighs around 49 kilodaltons. It is expressed prominently in the mitochondria of eukaryotic cells where it initiates the Krebs cycle by providing citrate to be further processed. Citrate synthetase is an important point of control within this metabolic cycle.
Biological function summary
Citrate synthetase plays a central role in energy production by converting oxaloacetate and acetyl-CoA into citrate. This enzyme is not known to be part of any larger complex but associates with downstream enzymes such as aconitase in the metabolic pathway. Its activity is essential for cellular respiration impacting the overall metabolic rate of the organism. The concentration of citrate produced serves as a checkpoint both for the continuation of the Krebs cycle and for feedback inhibition of glycolysis.
Pathways
Citrate synthetase is integral to the Krebs cycle and the related oxidative phosphorylation pathway. These pathways are essential for efficient ATP production. Citrate synthetase works closely with enzymes like isocitrate dehydrogenase and alpha-ketoglutarate dehydrogenase within the Krebs cycle. Through these interactions citrate synthetase ensures the proper flow of carbon through the cycle impacting ATP yield and cellular energy homeostasis.
Citrate synthetase's function can influence metabolic diseases like diabetes and mitochondrial disorders. Alterations in its activity may contribute to the dysregulation of glucose metabolism seen in diabetes affecting enzymes like glucose transporter 4 (GLUT4). In mitochondrial disorders changes in its normal activity can cause energy production deficiencies influencing proteins such as cytochrome c which is critical in the electron transport chain. Understanding these interactions can help develop therapeutic strategies targeting metabolic pathways.
