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描述 概述 Product Description
The active form of the enzyme is inhibited by citrate analogs, and fluoracetate. Other inhibitors include oxidative stress agents such as peroxynitrite, hydrogen peroxide and superoxide, which inactivate the enzyme by changing the [4Fe-4S] to a [3Fe-4S] cluster. Therefore, aconitase is considered a good marker of mitochondrial and cellular oxidative stress. This inhibition of mitochondrial aconitase can lead to a decreased energy production, whereas in cytosolic aconitase it triggers binding of the enzyme to mRNA iron response elements resulting in increased expression of iron uptake proteins and decreased transcription of iron sequestering protein.
A hydroxyl scavenging solution (Aconitase preservation solution) is supplied with this kit to maintain aconitase activity during sample preparation. An inactivated [3Fe-4FS] aconitase may be activated in vitro by the addition of iron and cysteine.Citrate ←→ cis-aconitate ←→ isocitrate
(↑ Abs. 240 nm)
The Aconitase Activity Assay Kit measures the activity of aconitase in a sample by following the conversion of isocitrate to cis-aconitate as indicated by an increase in absorbance at UV 240 nm with an extinction coefficient of 2.2 OD/mM per well.
Note: Mitochondrial and cytoplasmic aconitase activities are indistinguishable. Therefore, to measure the mitochondrial activity only, first isolate mitochondria, but to measure for both activities, separate the cells into cytoplasmic and mitochondrial fractions (see TB552 for details).
The entire assay can be completed within 2 hours. The intra-assay variation and inter-assay variation with this assay are each less than 10% and 15% respectively.
Panel Components
50 ml Buffer (Tube 1), (72575-50ML) 20 mlAconitase preservation solution, (72576-20ML) 800 µl 25X Isocitrate, (72577-800UL) 200 µl100X Manganese, (72578-200UL) 1 eaUV Microplate, (72579-1EA) 1 mlDetergent, (72580-1ML) * This kit contains sufficient materials for 96 measurements (one 96-well microplate).
Storage: Store all components at 2 to 8°C. Do not freeze.
Note: The 96-well microplate has a monoclonal antibody pre-bound to the wells. This plate can be separated into twelve 8 well strips and thus can be used for up to 12 individual experiments.产品目录编号 AAMT009 品牌系列 Novagen®
储存和货运信息 货运代码 Blue Ice Only
储存 +2°C to +8°C
无需冷冻 No
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