Biological Information
Background Information:
5-Lipoxygenase catalyzes the oxidative metabolism of arachidonic acid to 5-hydroxyeicosatetraenoic acid (5-HETE), the initial reaction leading to formation of leukotrienes. Leukotrienes (LTs) are inflammatory mediators causing, for example, phagocyte chemotaxis and increased vascular permeability. Inhibition of LT biosynthesis may be beneficial in inflammatory diseases.
Family:
Lipid Metabolism
Sub Family:
Eicosanoid Turnover
Class:
Lipoxygenase
Protein Name:
5-LO
Uniprot Number:
P09917
Protein Aliases:
Arachidonate 5-lipoxygenase B, 5-LO
Gene Name:
ALOX5
Gene ID:
240
Gene Aliases:
5-LOX
Accession Number:
NM_000698
Organism:
Human
Construct Details:
Full Length, Recombinant
Assay Information
Assay Type:
Biochemical
Assay Sub Type:
Enzymatic
Functional Mode:
Antagonist
Detection Method:
Spectrofluorimetry
Measured Response:
Fluorescence
Testing Information
Procedure Summary:
Human recombinant 5-LO expressed in insect Sf9 cells is used. Test compound or vehicle are preincubated with 1.4 U/mlǂ of enzyme for 15 minutes at 25°C in Tris buffer pH 7.4. The reaction is initiated by addition of 10 µM arachidonyl amide and dye DHR123 for another 20 minute incubation period. Read plate on a spectrophotometer (Excitation: 485 nm, Emission: 535 nm). Compounds are screened at 10 µM.
Substrate:
10 µM AMC Arachidonoyl Amide
Incubation:
20 min at 25°C
Control Inhibitor:
NDGA
Criteria For Significance:
≥ 50 of max stimulation or inhibition
Test Concentration / Dose:
Suggested concentration for initial testing is 10 uM
Minimum Order Quantity:
2
Turnaround Time
Standard:
15 Business Days
Reference Compound Data
Clinical Relevance
Therapeutic Area:
Inflammation/Allergy
Additional Information
Brand:
LeadHunter
Testing Location:
Taiwan - Taipei