Catalog # SP4631

Sphingosine Kinase 2 (Thr-578), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:500

Size 100 μl

Species Reactivity Hu, Rt, Ms

MW 70 kDa

Sphingolipids are metabolized into bioactive products that include ceramide, sphingosine, and sphingosine-1-phosphate (S1P). Sphingosine Kinase (SK) catalyzes the phosphorylation of the lipid sphingosine, creating S1P. S1P subsequently signals through cell surface G protein-coupled receptors, as well as intracellularly, to modulate cell proliferation, survival, motility and differentiation. Two isoforms of SK have been identified, SK1 and SK2. The mRNA for both of these isoforms is widely expressed with SK1 expression highest in brain, heart, kidney, thymus, spleen and lung, while SK2 is highest in kidney and liver. SKs can be activated through growth factor, G protein-coupled, and immunoglobulin receptor signalling. Regulation of SK1 and SK2 activity may occur through phosphorylation. SK1 is phosphorylated at Ser-225 by ERK leading to increased activity and translocation to the plasma membrane. SK2 is phosphorylated in response to EGF, PKC activators, and phorbol esters. ERK1 can phosphorylate both Ser-351 and Thr-578, and non-phosphorylatable mutants of these sites suppress ERK1-mediated chemotaxis.

References

Hait, N.C. et al. (2007) J Biol. Chem. 282(16):12058.
Hait, N.C. et al. (2005) J Biol. Chem. 280:29462.
Spiegel, S. & Milstien, S. (2003) Nat. Rev. Mol. Cell Biol. 4:397.

Western blot of human recombinant SK2 (lanes 1-4) and HeLa treated with calyculin A (lanes 5-8). The blots were untreated (lane 1, 3, 5 & 7) or treated with lambda phosphatase (lane 2, 4, 6 & 8), then probed with anti-SK2 (N-terminal region) (lanes 1, 2, 5 & 6) or anti-SK2 (Thr-578) (lanes 3, 4, 7, & 8).

Phospho-SK2 (Thr-578) synthetic peptide (coupled to KLH) corresponding to amino acid residues surrounding Thr-578 in human SK2. This peptide sequence is highly conserved in rat and mouse SK2 proteins, and is not well conserved in SK1.

*For more information, see UniProt Accession Q9NRA0

Rabbit polyclonal, affinity-purified antibody is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.

The products are are safely shipped at ambient temperature for both domestic and international shipments. Each product is guaranteed to match the specifications as indicated on the corresponding technical data sheet. Please store at -20C upon arrival for long term storage.

This antibody was affinity purified using phospho-SK2 (Thr-578) peptide (without carrier). The antibody detects 70 kDa* proteins corresponding to the molecular mass of SK2 on SDS-PAGE immunoblots of human recombinant SK2 and endogenous SK2 in human HeLa cells treated with calyculin A. This reactivity is not observed after lambda phosphatase treatment.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.

Product References:

Xia, C. et al. (2018) Antiviral Res. 158:171. (WB: human A549 cells)
Fu, P. et al. (2016) J Biol Chem. 291(53):27187. (WB: human endothelial cells)
Wang, X. et al. (2016) PLoS Pathog. 12(10):e1005926. (WB: brain microvascular endothial cells)
Wallington, C.T. et al. (2014) Cancer Res(74):2803. (WB: human leukemia cells)
Dai, L. et al. (2013) Mol Endocrin 28(2):197. (WB: MCF7)

This kit contains: