Citrate Synthase Activity Assay Kit (ab119692)
Key features and details
- Assay type: Enzyme activity
- Detection method: Colorimetric
- Platform: Microplate reader
- Assay time: 3 hr 15 min
- Sample type: Cell culture extracts, Tissue Extracts
- Sensitivity: 1.56 µg/ml
Overview
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Product name
Citrate Synthase Activity Assay Kit
See all Citrate synthetase kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% 1 3 5.45% Inter-assay Sample n Mean SD CV% 1 3 8.3% -
Sample type
Cell culture extracts, Tissue Extracts -
Assay type
Enzyme activity -
Sensitivity
1.56 µg/ml -
Range
1.56 µg/ml - 100 µg/ml -
Assay time
3h 15m -
Species reactivity
Reacts with: Cow, Human
Does not react with: Mouse, Rat -
Product overview
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 -
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.
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. -
Platform
Microplate reader
Properties
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Storage instructions
Please 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 (ab260490) 1 x 15ml -
Research areas
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Pathway
Carbohydrate metabolism; tricarboxylic acid cycle; isocitrate from oxaloacetate: step 1/2. -
Sequence similarities
Belongs to the citrate synthase family. -
Cellular localization
Mitochondrion matrix. - Information by UniProt
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Alternative names
- CISY_HUMAN
- Citrate synthase
- Citrate synthase, mitochondrial
see all -
Database links
- Entrez Gene: 280682 Cow
- Entrez Gene: 1431 Human
- Omim: 118950 Human
- SwissProt: Q29RK1 Cow
- SwissProt: O75390 Human
- Unigene: 430606 Human
- Unigene: 728168 Human
Images
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ELISA - Citrate Synthase Activity Assay Kit (ab119692)Figure 2. Example activity measurements from titrated of Human heart homogenate.
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ELISA - Citrate Synthase Activity Assay Kit (ab119692)Figure 1. Example activity measurements from titrated HepG2 cells.
