Detection of Phospho-5‑Lipoxygenase (S523) by Western Blot. Western blot of rat brain (cortex) tissue lysate showing specific immunolabeling of the ~80 kDa doublet of 5-LO phosphorylated at S523 (Control). The phosphospecificity of this labeling is demonstrated by treatment with 1200 U of lambda Phosphatase ( lambda -PPase) for 30 minutes before being exposed to the Anti-Phospho-5-Lipoxygenase (S523). The immunolabeling is completely eliminated by treatment with lambda -PPase.
Preparation and Storage
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
For long-term storage, ≤ -20° C is recommended. Product is stable at ≤ -20° C for at least 1 year.
5-Lipoxygenase (5-LO) is an 80 kDa, 676 amino acid (aa) nonheme iron
dioxygenase. It converts nuclear membrane arachidonic acid into
leukotriene A4 (LTA4) that is then modified to
produce either LTB4 or
LTC4. 5-LO is found in leukocytes and generally
exists in the cytoplasm. Upon activation, 5-LO will either remain
cytosolic or undergo translocation into the nucleus. In the nucleus, its
enzyme products impact gene expression; in the cytosol, its products
are secreted and contribute to inflammation. 5-LO has three Nuclear
Localization Signal (NLS) motifs starting at amino acids 112, 158, and
518. Upon cell activation, Ser at 523 is phosphorylated by Protein
Kinase A (PKA). This amino acid lies within the principal NLS of 5-LO.
When phosphorylated, the NLS is inactivated, blocking 5-LO entry into
the nucleus. This appears to impact overall leukotriene biosynthesis,
and potentially serves as a regulatory mechanism for inflammation.
Jones, S.M. et al. (2003) J. Biol. Chem. 278:10257.
Dixon, R.A. et al. (1988) Proc. Natl. Acad. Sci. USA 85:416.
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