Description
General description
Recombinant, human 5-Lipoxygenase expressed in S. fruigiperda insect cells using a baculovirus overexpression system. 5-LO catalyzes the formation of 5(S)-HpETE from arachidonic acid and its subsequent conversion to leukotriene A₄. It is localized in the cytosol of resting human and rat peripheral blood neutrophils, whereas it is found in the nucleus in rat basophilic leukemia cells and human alveolar macrophages. Upon stimulation of the cell, 5-LO translocates to the nuclear membrane where it is found in association with 5-LO activating protein (FLAP) and with the 85 kDa cPLA₂.
Physical form
In 100 mM Tris containing 5 mM EGTA, pH 8.0.
Preparation Note
Following initial thaw, aliquot and freeze (-70°C).
Other Notes
Brock, T.G., et al. 1997. J. Biol. Chem.272, 8276.
Pouliot, M., et al. 1996. Eur. J. Biochem.238, 250.
Zhang, Y.Y., et al. 1993. J. Biol. Chem.268, 2535.
Shimizu, T., et al. 1984. Proc. Natl. Acad. Sci. USA81, 689.
One unit is defined as the amount of enzyme that consumes 1.0 nmol of oxygen per min at 25°C in 50 mM Tris-HCl, 100 µM arachidonate, 2 mM CaCl₂, 1 mM ATP, 250 µM oxygen. Oxygen consumption can be measured using a oxygraph equipped with a Clark oxygen electrode.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Standard Handling (A)