Catalog # SK6590

Sphingosine Kinase 2 Phospho-Regulation Antibody Kit

Antibody Sampler Kit

DATASHEET  




$395

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
Spiegel, S. & Milstien, S. (2003) Nat. Rev. Mol. Cell Biol. 4:397.
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

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

Western blot of HeLa treated 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-SK2 (N-terminal region) (lanes 1 & 2) or anti-SK2 (Thr-578) (lanes 3 & 4).



Rabbit polyclonal antibodies are supplied in phosphate-buffered saline (PBS), 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. The secondary reagents are supplied in PBS without azide, and the peptides are supplied in PBS and 0.05% sodium azide. Store all 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.



*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:

SP4621 Wang, X. et al. (2016) PLoS Pathog. 12(10):e1005926. WB: Brain endothelial cells
SP4631 Wang, X. et al. (2016) PLoS Pathog. 12(10):e1005926. WB: Brain endothelial cells
SP4621 Neubauer, H & Pitson, SM (2016) F1000Research. 5:2825. IP-WB, ICC: HEK293, HeLa, MEF
SP4621 Brunno, G et al. (2015) BBA MCB Lipids. 1851.2:194. WB: murine myoblast
SP4621 St Patrick, R. et al. (2015) Emerg micro & infect. 4(10):e61 ICC: hSKMC, MCF10A
SP4621 Wallington-Beddoe, CT et al. (2014) Cancer Res 74(10):2803. WB: ALL cell lines, K562 siRNA KD
SP4631 Wallington-Beddoe, CT et al. (2014) Cancer Res 74(10):2803. WB: ALL cell lines
SP4631 Dai, L. et al. (2013) Molec Endocrinology 28(2):197. WB: MCF7 + Insulin
RS3251 Kawasaki, H. et al. (2013) World J Gastroenter. 19(17):2629. WB, ICC: mouse intestinal myofibroblasts
RS3251 Estrada-Bernal, A. et al. (2011) J Neurooncol. 102:353. Western Blot

This kit contains:

CATALOG#DESCRIPTIONSIZEAPPLICATIONSSPECIES REACTIVITYMW (kDa)
Sphingosine Kinase 2 (N-terminal region) Rabbit pAb50 μlWB, E, IP, ICCHu, Rt, Ms70
Sphingosine Kinase 2 (Thr-578), phospho-specific Rabbit pAb50 μlWB, EHu, Rt, Ms70
Sphingosine Kinase 2 (N-terminal region) Peptide50 μgAB, E
phospho-Sphingosine Kinase 2 (Thr-578) Peptide 50 μgAB, E
Anti-Rabbit Ig Light-Chain Specific:HRP Mouse mAb100 μlWB, E, ICC, IHCRb
KIT SUMMARY

The Sphingosine Kinase 2 (SK2) phospho-regulation antibody kit can be used to detect phosphorylation of Thr-578 relative to total SK2 expression. The kit includes rabbit polyclonal antibodies, blocking peptides, and secondary reagents for detection in western blot.