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MAK051

Aconitase Activity Assay Kit

sufficient for 100 colorimetric tests

Synonym(e):

Aconitase Enzyme Activity Kit

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Über diesen Artikel

NACRES:
NA.84
UNSPSC Code:
12161503


usage

sufficient for 100 colorimetric tests

detection method

colorimetric

relevant disease(s)

neurological disorders; cardiovascular diseases; cancer

storage temp.

2-8°C

Gene Information

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

Aconitase Activity Assay Kit has been used to measure the activity of cytosolic and mitochondrial aconitase enzyme.
Suitable for the measurement of aconitase activity in biological samples including tissue and cells

Biochem/physiol Actions

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.

Features and Benefits

Compatible with high-throughput handling systems.


pictograms

Health hazardCorrosion

signalword

Danger

hcodes

Hazard Classifications

Eye Dam. 1 - Resp. Sens. 1 - Skin Corr. 1B

Lagerklasse

8A - Combustible corrosive hazardous materials



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Questions

  1. 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
    1. 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.

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