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Technical Data Sheet

SM5401

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Sphingosine Kinase 1 (C-terminal region)
Rat Monoclonal
Price
Size
Species Reactivity
MW

$295
100 μl
Hu, Rt, Ms
47 kDa

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Enlarge

Western blot image of human Hela cells (lanes 1 & 2) and recombinant his-tagged human SK1 protein (lanes 3 & 4). The blots were probed with rat monoclonal anti-SK1 (C-terminal Region) at 1:250 (lanes 1 & 3) and 1:1000 (lanes 2 & 4).

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Enlarge

Immunocytochemical labeling of sphingosine kinase 1 (SPHK1) in paraformaldehyde fixed and NP-40 permeabilized A431 cells. The cells were labeled with rat monoclonal anti-SPHK1 (SM5401). The antibody was detected using goat anti-rat DyLight® 594.

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Application
Dilution


ELISA
1:1000


ICC
1:200


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.
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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. 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.


Background References
Melendez, A.J. et al. (2000) Gene 251(1) :19.
Pitson, S.M. et al. (2003) EMBOJ. 22(20) :5491.
Pitson, S.M. et al. (2005) J Exp. Med. 201(1) :49.
Immunogen
Clone M540 was generated from a SK1 synthetic peptide (coupled to carrier) corresponding to amino acids in the C-terminal region of mouse SK1. This sequence has high homology to human and rat SK1, and is not homologous to sequences in SK2.
Buffer and Storage
Rat monoclonal, 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 SK1 (C-terminal Region) peptide. The purified antibody detects 42 kDa* full-length recombinant human and mouse SK1 proteins, as well as detects 42, 50, and 60 kDa forms of SK1 in human HeLa cells.

*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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