QuantiChrom™ Citrate Synthase Assay Kit

Protocol SDS

Application

• Direct assays of citrate synthase activity in cell lysates, tissues, and other biological samples.

Key Features

• Sensitive and accurate. Linear detection range 0.50 to 120 U/L citrate synthase activity in 96-well plate assay.
• Simple and convenient. The procedure involves adding a single working reagent and reading the optical density at 5 min and 10 min at room temperature.
• High-throughput. Can be readily automated as a high-throughput 96-well plate assay for thousands of samples per day.

Method

• OD412nm

Samples

• Cell lysates, tissues, and other biological samples

Species

• All

Procedures

• 10 min

Size

• 100 tests in 96-well plate

Detection Limit

• 0.50 U/L

Shelf Life

• 6 months

More Details

CITRATE SYNTHASE (E.C. 2.3.3.1) is a pivotal enzyme that initiates the tricarboxylic acid (TCA) cycle. It catalyzes the condensation reaction between acetyl-CoA and oxaloacetic acid, yielding citric acid. Citrate synthase levels of cells and tissue are used as a guide in determining mitochondrial dysfunctions and cellular metabolism. High levels of citrate synthase activity indicate increased metabolic activity or cellular energy production. This can occur in situations where cells are actively metabolizing substrates for energy production, such as during periods of increased physical activity or in tissues with high energy demands, like muscle tissue. Conversely, low levels of citrate synthase activity may suggest reduced metabolic activity or energy production. This could be seen in situations where cellular metabolism is slowed down, such as during periods of rest or in conditions associated with mitochondrial dysfunction. Simple, direct and automation-ready procedures for measuring citrate synthase activity are very desirable. BioAssay Systems’ QuantiChrom™ Citrate Synthase Assay is based on an improved Ellman method, in which coenzyme A thiol produced by the action of citrate synthase forms a yellow color with 5,5′-dithiobis (2-nitrobenzoic acid). The intensity of the product color, measured at 412 nm, is proportionate to the enzyme activity in the sample.

Does this kit supply Citrate Synthase?

No, this kit is for the measurement of Citrate Synthase activity, so researchers must supply their own Citrate Synthase enzyme or sample specific to their needs.

What is the species origin of the Citrate Synthase used to develop this kit?

Citrate Synthase from E. coli was used to develop the kit, but the kit has been used to detect Citrate Synthase in variety of cell lines and tissue homogenates.

How long are the solubilized reagents good for?

Reagent A and Reagent B should be used within two (2) weeks of dissolution.

For more detailed product information and questions, please feel free to email or call us at 1-510-782-9988 x 2. For more general information regarding our assays, please refer to Technical Support.

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If you or your labs do not have the equipment or scientists necessary to run this assay, BioAssay Systems can perform the service for you.

– Fast turnaround
– Quality data
– Low cost

Please email or call 1-510-782-9988 x 2 to discuss your projects.