Catalog # SP1641

Sphingosine Kinase 1 (Ser-225), phospho-specific Antibody

Rabbit Polyclonal

Application / Dilution

ELISA	1:2000
WB	1:1000

Size 100 μl

Species Reactivity Hu

MW 40-50 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. SK1 has been shown to mediate cell growth, prevention of apoptosis, and cellular transformation, and is upregulated in a variety of human tumors. Regulation of SK1 may occur through ERK mediated phosphorylation of Ser-225. This phosphorylation leads to increased activity and translocation to the plasma membrane.

References

Pitson, S.M. et al. (2005) J Exp. Med. 201(1) :49.
Pitson, S.M. et al. (2003) EMBOJ. 22(20) :5491.
Melendez, A.J. et al. (2000) Gene 251(1) :19.

Western blot image of recombinant his-tagged human SK1 protein that was phosphorylated with ERK-2. Blots were probed with anti-SK1 (Central Region) (SP1621; lanes 1-3) and anti-SK1 (Ser-225) (SP1641; lanes 4-6). Both antibodies were used in the presence of no peptide (lanes 1 & 4), phospho-SK1 (Ser-225) peptide (SX1645; lanes 2 & 5), or unphosphorylated SK1 (Ser-225) peptide (SX1625; lanes 3 & 6).

Western blot of HeLa stimulated with calyculin A (lanes 1-4). The blots were untreated (lane 1 & 3) or treated with lambda phosphatase (lane 2 & 4), then probed with anti-SK1 (Central region) SP1621 (lanes 1 & 2) or anti-SK1 (Ser-225) SP1641 (lanes 3 & 4).

Phospho-SK1 (Ser-225) synthetic peptide (coupled to carrier protein) corresponding to amino acids surrounding serine 225 in human SK1. This sequence has 4 amino acid differences from mouse and 5 from rat SK1, and is not homologous to sequences in SK2. The conserved site in rat and mouse SK1 is at serine 224.

*For more information, see UniProt Accession Q9NYA1

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 cross-adsorbed to non-specific phosphoserine and unphosphorylated SK1 (Ser-225) peptide before affinity purification using phospho-SK1 (Ser-225) peptide. The purified antibody detects a 47 kDa* full-length recombinant human SK1 protein phosphorylated with ERK2.

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

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