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For enzymatic assays, it is advised to scrape cells with a cell scraper in assay buffer, pellet them, and then use a specific volume of assay buffer to lyse the cells. Alternatively, trypsin can be used. However, after pelleting the cells post-trypsinization, it is important to wash them 2 times in 1x PBS to remove any trace amounts of trypsin.
MAK051
Aconitase Activity Assay Kit
sufficient for 100 colorimetric tests
Synonym(s):
Aconitase Enzyme Activity Kit
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About This Item
UNSPSC Code:
12161503
NACRES:
NA.84
Recommended Products
usage
sufficient for 100 colorimetric tests
detection method
colorimetric
relevant disease(s)
neurological disorders; cardiovascular diseases; cancer
storage temp.
2-8°C
Gene Information
human ... ACO1(48) , ACO2(50)
mouse ... ACO1(11428) , ACO2(11429)
rat ... ACO1(50655) , ACO2(79250)
General description
The aconitases are a family of iron-sulfur containing enzymes that catalyzes the isomerization of citrate to isocitrate. Two aconitases, encoded by different genes, have been identified in eukaryotes. Aconitase 1 (ACO1, c-aconitase) is a cytosolic enzyme while Aconitase 2 (ACO2, m-aconitase) is a mitochondrial enzyme that functions in the tricarboxylic acid cycle. Aconitases are reversibly inactivated by oxidative stress, and aconitase activity in cells and tissues has been used as a biomarker for oxidative damage.
Application
Features and Benefits
Compatible with high-throughput handling systems.
Suitability
Suitable for the measurement of aconitase activity in biological samples including tissue and cells
Principle
The Aconitase Activity Assay kit provides a simple and direct procedure for measuring Aconitase activity in a variety of samples. Aconitase activity is determined in a coupled enzyme reaction in which citrate is converted to isocitrate by aconitase. This results in a colorimetric (450 nm) product proportional to the enzymatic activity present. One unit of aconitase is the amount of enzyme that will isomerize 1.0 μmole of citrate from isocitrate per minute at pH 7.4 at 25 °C.
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Danger
hcodes
Hazard Classifications
Eye Dam. 1 - Resp. Sens. 1 - Skin Corr. 1B
Storage Class
8A - Combustible corrosive hazardous materials
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What is the best method to lift adherent cells from the plate for the aconitase activity assay kit MAK051? Is using the included assay buffer and a cell scraper sufficient, or would you recommend washing with a detergent first? The tech bulletin mentions the use of 10E6 (suspended) cells, but it is not clear how to prepare adherent cells for use with the kit.
1 answer-
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