Product nameCitrate Synthase Activity Assay Kit
Intra-assay Sample n Mean SD CV% 1 3 5.45% Inter-assay Sample n Mean SD CV% 1 3 8.3%
Sample typeCell culture extracts, Tissue Extracts
Assay typeEnzyme activity
Range1.56 µg/ml - 100 µg/ml
Assay time3h 15m
Species reactivityReacts with: Human
Does not react with: Mouse, Rat
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
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 126.96.36.199). This enzyme is an exclusive marker of the mitochondrial matrix. Citrate synthase is found in nearly all cells capable of oxidative metabolism.
Storage: All components are shipped cold. 10X Reaction Reagent mixture is shipped lyophilized. Before use, rehydrate the reaction mixture by adding 1mL base buffer. Vortex the tube thoroughly to dissolve the lyophilized powder to prepare 10X Reaction solution. After hydration unused amounts of the 10X Reaction solution should be stored at -80°C for 6 months. Store all other components at 4°C. This kit is stable for 6 months from receipt.
Storage instructionsPlease refer to protocols.
Components 1 x 96 tests 10X Blocking Buffer 1 x 6ml 10X Reaction Reagent 1 vial 1X Base Buffer 1 x 25ml 20X Buffer 1 x 20ml CS Microplate 1 unit Extraction Buffer 1 x 15ml
- Pathways and Processes
- Metabolic signaling pathways
- Energy transfer pathways
- Energy Metabolism
PathwayCarbohydrate metabolism; tricarboxylic acid cycle; isocitrate from oxaloacetate: step 1/2.
Sequence similaritiesBelongs to the citrate synthase family.
Cellular localizationMitochondrion matrix.
- Information by UniProt
- Citrate synthase
- Citrate synthase, mitochondrial
This product has been referenced in:
- Zhao Z & Pu Y Lixisenatide enhances mitochondrial biogenesis and function through regulating the CREB/PGC-1a pathway. Biochem Biophys Res Commun 508:1120-1125 (2019). Read more (PubMed: 30553453) »
- Corbet C et al. Interruption of lactate uptake by inhibiting mitochondrial pyruvate transport unravels direct antitumor and radiosensitizing effects. Nat Commun 9:1208 (2018). Read more (PubMed: 29572438) »