Description
Key facts
Detection method:Colorimetric,
Sample types:Cell culture extracts, Tissue Extracts,
Reacts with:Cow, Human,
Assay type:Enzyme activity,
Sensitivity:= 1.56 µg/mL,
Range:1.56 - 100 µg/mL,
Assay time:3h 15m,
Assay Platform:Microplate reader
Product details:
Citrate Synthase Activity Assay Kit (ab119692) is used to determine mitochondrial citrate synthase (CS) activity in a sample. The enzyme is immunocaptured within the wells of a microplate, and activity is determined by recording color development of TNB, which is generated in the reaction of citrate synthesis. The reaction product, TNB, absorbs at OD = 412 nm.
This kit immunocaptures only native citrate synthase from the samples. Using the immunocapture method allows the measurement of citrate synthase activity with simple sample preparation and no need for mitochondrial isolation.
This kit can be used with whole cell extracts or tissue homogenates.
Citrate synthase assay protocol summary
- add samples to wells
- incubate for 3 hrs to capture enzyme
- wash with wash buffer
- add activity solution
- analyze with microplate reader for 5-30 min at 20s intervals
Related and recommended products
For a citrate synthase assay without the antibody pulldown step, see Citrate synthase assay kit
ab239712
Other Notes
Citrate synthase (CS, O75390) is the initial enzyme of the tricarboxylic acid (TCA) cycle. This enzyme is a 51.7 kDa enzyme that catalyzes the reaction of 2 carbon acetyl CoA with 4 carbon oxaloacetate to form the 6 carbon citrate (EC 2.3.3.1). This enzyme is an exclusive marker of the mitochondrial matrix. Citrate synthase is found in nearly all cells capable of oxidative metabolism.
Properties and Storage Information:
Shipped at conditions-Blue Ice, Appropriate short-term storage conditions-Multi, Appropriate long-term storage conditions-Multi, Storage information-Please refer to protocols
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.
