Exalpha Biologicals, Inc.

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FIX&PERM Cell Fixation and Permeabilization Kit

Flow cytometric analyses with monoclonal antibodies were so far mainly restricted to cell surface molecules. Intracellular structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, immunoglobulins etc. were largely excluded from such studies. Also excluded from flow cytometric studies were cytoplasmic localizations of well-established membrane molecules like CD3 and CD22, which, in their cytoplasmic form, are the most reliable lineage markers in undifferentiated leukemia. With the FIX&PERM® Kit flow cytometric analysis of intracellular antigens has become as easy as surface antigen studies. The only prerequisite is the availability of suitable antibody conjugates. Most of the available monoclonal antibody conjugates can be used with the FIX&PERM® Kit, some determinants are sensitive, however, to the fixation step involved. This and the optimal fixation time have to be tested for each reagent.

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Exalpha Biologicals, Inc.

Sphingosine Kinase 1 (Full Length) with N-Terminal HIS tag

  • Product Code: X1708E
  • Size: 10 µg
  • Availability: In Stock In Stock
  • Price (USD): $361

Cat #

X1708E		 Quantity:      

Data Sheet

Product Name

Sphingosine Kinase 1 (Full Length) with N-Terminal HIS tag

Host/Source

Sf9 cells

Isotype

N/A

Product Type

Active Enzyme

Reactivity

Human

Applications

Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.

Size

10 µg

Price (USD)

$361

Background

Sphingosine Kinase 1 (Sphk1) catalyzes the phosphorylation of sphingosine to sphingosine 1-phosphate (S1P), an important signaling molecule with intra- and extracellular functions. Inside the cell S1P acts as a signaling molecule like other sphingolipid metabolites like ceramide and sphingosine. S1P has been implicated in regulating cell differentiation, calcium mobilization from intracellular stores, and apoptosis. The cell surface receptors for S1P are the EDG family of G protein-coupled receptors (S1P Receptors). These receptors couple to multiple G proteins (e.g. S1P1 couples to Gi whereas S1P2 and S1P3 couple to Gq, G13 in addition to Gi) and regulate a extremely wide range of cellular events including cell motility, survival, apoptosis, migration and cell-cell interaction. Important roles for S1P have also been reported in regulation of cardiogenesis, vascular maturation, oocyte survival, immune cell trafficking, cells of the neuronal system and bone cells. S1P levels are regulated by the activity of Sphk (Sphk1 and Sphk2).

Immunogen

Baculovirus infected Sf9 cells

Formulation

Provided in 25 mM Tris-HCl, 100 mM NaCl, pH 8.0, 0.05% Tween, 3 mM DTT and 50% glycerol

Customer Storage

Enzyme should be stored at -20°C. Enzyme should be kept on ice when dispensing

Target Molecular Weight

46.9 kDa

Database Links:

SwissProtAF200328Human

References

1. Kihara A, Ikeda M, Kariya Y, Lee EY, Lee YM, Igarashi Y, Sphingosine-1-phosphate lyase is involved in the differentiation of F9 embryonal carcinoma cells to primitive endoderm. J. Biol. Chem. 278 (2003) 14578-14585

2. Choi OH, Kim JH, Kinet JP, Calcium mobilization via sphingosine kinase in signalling by the FceRI antigen receptor. Nature 380 (1996) 634-636

3. Olivera A, Kohama T, Edsall L, Nava V, Cuvillier O, Poulton S, Spiegel S, Sphingosine kinase expression increases intracellular sphingosine-1-phosphate and promotes cell growth and survival. J. Cell Biol. 147 (1999) 545-558

4. Chun J, Goetzl EJ, Hla T, Igarashi Y, Lynch KR, Moolenaar W, Pyne S, Tigyi G, International union of pharacology. XXXIV. Lysophospholipid receptor nomenclature. Pharmacol. Rev. 54 (2002) 265-269

5. Hla T, Signaling and biological actions of sphingosine 1-phosphate. Pharmacol. Res. 47 (2003) 401-407

6. Gregorius K, Mouritsen S, Elsner HI, Hydrocoating: a new method for coupling biomolecules to solid phases. J. Immunol. Methods, 181 (1995) 65-73

7. Zhang JH, Chung TDY, Oldenburg KR, A simple statistical parameter for use in evaluation and validation of high throughput screening assays. J. Biomol. Screening, 4 (1999) 67-73

8. Olivera, A., Kohama, T., Tu, Z., Milstien, S., and Spiegel, S. (1998) J. Biol. Chem. 273, 12576?12583

9. Zhang, H., Desai, N. N., Olivera, A., Seki, T., Brooker, G., and Spiegel, (1991) J. Cell Biol. 114, 155?167

10. Olivera, A., Rosenthal, J., and Spiegel, S. (1996) J. Cell. Biochem. 60, 529?537

11. Zhang, H., Desai, N. N., Olivera, A., Seki, T., Brooker, G., and Spiegel, S. (1991) J. Cell Biol. 114, 155?167

12. Ohotski, J., et al. Expression of sphingosine 1-phosphate receptor 4 and sphingosine kinase 1 is associated with outcome in oestrogen receptor-negative breast cancer. Br. J. Can. (2012), 106, 1453-1459