Lipoxygenase Assay Kit (ab241038)
Key features and details
- Detection method: Fluorescent
- Sample type: Cell Lysate, Other biological fluids, Serum, Tissue Lysate
Overview
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Product name
Lipoxygenase Assay Kit -
Detection method
Fluorescent -
Sample type
Serum, Other biological fluids, Cell Lysate, Tissue Lysate -
Species reactivity
Reacts with: Mammals -
Product overview
In the Lipoxygenase Assay Kit (ab241038) lipoxygenase converts the LOX substrate to an intermediate that reacts with the probe generating a fluorescent product. The increase in fluorescent signal can be recorded at Ex/Em 500/536 nm and is directly proportional to LOX activity.
The kit includes 5-lipoxygenase enzyme (LOX Enzyme) as a positive control. A lipoxygenase inhibitor that completely inhibits lipoxygenase activity is also included to calculate the specific activity of LOX in biological samples.
The kit can detect as low as 0.004 mU/mg protein.
Properties
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Storage instructions
Store at -20°C. Please refer to protocols. -
Components 100 tests LOX Assay Buffer 1 x 25ml LOX Enzyme 1 x 20µl LOX Inhibitor 1 x 100µl LOX Lysis Buffer 1 x 2ml LOX Probe 1 x 200µl LOX Substrate 1 x 6µl Oxidized Probe Standard 1 x 200µl -
Research areas
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Relevance
The gene for arachidonate 5-lipoxygenase encodes a member of the lipoxygenase gene family and plays a dual role in the synthesis of leukotrienes from arachidonic acid. The encoded protein, which is expressed specifically in bone marrow-derived cells, catalyzes the conversion of arachidonic acid to 5(S)-hydroperoxy-6-trans-8,11,14-cis-eicosatetraenoic acid, and further to the allylic epoxide 5(S)-trans-7,9-trans-11,14-cis-eicosatetrenoic acid (leukotriene A4). Leukotrienes are important mediators of a number of inflammatory and allergic conditions. Mutations in the promoter region of this gene lead to a diminished response to antileukotriene drugs used in the treatment of asthma and may also be associated with atherosclerosis and several cancers. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. The gene for arachidonate 12-lipoxygenase, 12R type encodes an enzyme involved in the converstion of arachidonic acid to 12R-hydroxyeicosatetraenoic acid. Mutations in this gene are associated with nonbullous congenital ichthyosiform erythroderma. The gene for arachidonate 15-lipoxygenase, type B encodes a member of the lipoxygenase family of structurally related nonheme iron dioxygenases involved in the production of fatty acid hydroperoxides. The encoded protein converts arachidonic acid exclusively to 15S-hydroperoxyeicosatetraenoic acid, while metabolizing linoleic acid less effectively. This gene is located in a cluster of related genes and a pseudogene that spans approximately 100 kilobases on the short arm of chromosome 17. Alternatively spliced transcript variants encoding different isoforms have been described. -
Cellular localization
Arachidonate 15-lipoxygenase B: Cytoplasm Arachidonate 5-lipoxygenase: Cytoplasm. Nucleus matrix. Nucleus membrane; Peripheral membrane protein. Note: Shuttles between cytoplasm and nucleus. Found exclusively in the nucleus, when phosphorylated on Ser-272. Calcium binding promotes translocation from the cytosol and the nuclear matrix to the nuclear envelope and membrane association. -
Alternative names
- 12R-lipoxygenase
- 12R-LOX
- 15-lipoxygenase 2
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Images
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Oxidized probe standard curve.Oxidized probe standard curve.
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Lipoxygenase enzyme reaction in positive control and in MCF-7 lysate.Lipoxygenase enzyme reaction in positive control and in MCF-7 lysate (10 µg protein).
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LOX activity in positive control and MCF-7 lysate.LOX activity in positive control and MCF-7 lysate. Assays were performed following the kit protocol.