Description
Key facts
Detection method:Colorimetric/Fluorometric,
Sample types:Urine, Plasma, Tissue Extracts, Cell culture supernatant, Serum,
Assay type:Quantitative,
Sensitivity:> 0.002 mM,
Range:0.002 - 10 mM,
Assay time:40m,
Assay Platform:Microplate reader
Product details:
Citrate Assay Kit ab83396 provides a simple, sensitive and rapid means of quantifying citrate in biological samples.
In the citrate assay protocol, citrate is converted to pyruvate via oxaloacetate. The pyruvate is quantified by converting a nearly colorless probe to an intensely colored (570 nm) and fluorescent (Ex/Em, 535/587 nm) product.
The citrate assay kit can detect 0.1 to 10 nmoles (~2 uM-10 mM) of citrate.
Citrate assay protocol summary:
- add samples and standards to wells
- add reaction mix and incubate for 30 min at room temp
- analyze with microplate reader
Other Notes
This product was previously called K655 Citrate Colorimetric/Fluorometric Assay Kit. Biovision was acquired by Abcam in 2021.
Citric acid (HOOC-CH
-C(-OH)(-COOH)-CH
-COOH) is a key intermediate in the TCA cycle which occurs in mitochondria. It is formed by the addition of oxaloacetate to the acetyl group of acetyl-CoA derived from the glycolytic pathway. Citrate can be transported out of mitochondria and converted back to acetyl CoA for fatty acid synthesis.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
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 also known as citric acid is a tricarboxylic acid with the molecular mass of approximately 192.13 g/mol. It is a small molecule that plays a critical role in metabolism. Citrate exists extensively in the mitochondria of eukaryotic cells and in the cytosol. Its high abundance in these locations allows it to participate in various biochemical reactions necessary for cellular energy production. Citrate forms through the condensation of acetyl-CoA and oxaloacetate by the enzyme citrate synthase initiating the tricarboxylic acid (TCA) cycle also known as the Krebs cycle.
Biological function summary
The presence of citrate is essential in the regulation of the TCA cycle where it functions as an important intermediate. Among its functions citrate can inhibit the enzyme phosphofructokinase-1 linking glycolysis and the TCA cycle. This inhibition ensures a balance between the supply of glucose and the demand for ATP. Citrate does not form part of a complex but it interacts with various metabolic enzymes establishing its role as a metabolic regulator. Additionally citrate acts as a substrate for ATP-citrate lyase aiding in the production of acetyl-CoA in the cytosol vital for fatty acid synthesis.
Pathways
Citrate involves itself significantly in the TCA cycle and fatty acid synthesis. The TCA cycle a central component of cellular respiration generates reducing equivalents for oxidative phosphorylation by processing acetyl-CoA. As citrate is part of this pathway it interacts with enzymes such as aconitase and isocitrate dehydrogenase. In fatty acid synthesis citrate serves as an acetyl-CoA donor after being converted by ATP-citrate lyase. This conversion links citrate to important pathways such as lipogenesis and cholesterol synthesis highlighting its diverse roles in cellular metabolism.
Citrate has a significant relationship with conditions like cancer and metabolic syndrome. Increased citrate production or its accumulation can occur in cancer cells as tumors often exhibit altered metabolism known as the Warburg effect. In these cases regulatory proteins like hypoxia-inducible factor (HIF) can affect citrate metabolism. In metabolic syndrome altered citrate levels can contribute to disrupted metabolic pathways impacting proteins such as acetyl-CoA carboxylase and fatty acid synthase. Understanding citrate's function in these diseases potentially offers insights into therapeutic targets.
