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Technical Data Sheet

SP4621

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Sphingosine Kinase 2 (N-terminal region)
Rabbit Polyclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
70 kDa

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Enlarge

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).

Application
Dilution


ELISA
1:2000


ICC
1:100


IP
1:100


WB
1:250



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
Product References
Neubauer, H & Pitson, SM (2016) F1000Research. 5:2825. IP-WB, ICC: HEK293, HeLa, MEF
Wang, X. et al. (2016) PLoS Pathog. 12(10):e1005926. WB: Brain endothelial cells
Brunno, G et al. (2015) BBA MCB Lipids. 1851.2:194. WB: murine myoblast
St Patrick, R. et al. (2015) Emerg micro & infect. 4(10):e61 ICC: hSKMC, MCF10A
Wallington-Beddoe, CT et al. (2014) Cancer Res 74(10):2803. WB: ALL cell lines, K562 siRNA KD
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Background
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.


Background References
Spiegel, S. & Milstien, S. (2003) Nat. Rev. Mol. Cell Biol. 4:397.
Hait, N.C. et al. (2005) J Biol. Chem. 280:29462.
Hait, N.C. et al. (2007) J Biol. Chem. 282(16):12058.
Immunogen
SK2 (N-terminal region) synthetic peptide (coupled to KLH) corresponding to amino acid residues in the N-terminal region of human SK2. This peptide sequence is highly conserved in rat and mouse SK2 proteins, and has no homology to SK1.
Buffer and Storage
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.
Specificity
This antibody was affinity purified using SK2 (N-terminal region) 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. The antibody also works for ELISA, immunoprecipitation, and immunocytochemistry.

*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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