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.

Ceramide Kinase (CERK)

  • Product Code: X2076E
  • Size: 10 µg
  • Availability: In Stock In Stock
  • Price (USD): $361

Cat #

X2076E		 Quantity:      

Data Sheet

Product Name

Ceramide Kinase (CERK)

Synonyms

Ceramide kinase, Acylsphingosine kinase, hCERK, Lipid kinase 4, LK4,

Host/Source

Sf9 cells

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

Sphingolipids, in addition to being structural components of membranes, regulate cell-cell and cell-substrate interactions, proliferation, and differentiation. Members of this diverse group of lipids have emerged as a novel class of signaling molecules that also regulate phagocytosis. The mechanisms by which sphingolipids exert these effects remain incompletely defined. More than a decade ago, it was found that ceramide can be phosphorylated to ceramide 1-phosphate (C1P). Ceramide kinase (CERK) and its phosphorylated product ceramide 1-phosphate (C1P) are central players in inflammation and cancer. The product of CERK activity, ceramide 1-phosphate (C1P), has been reported to have mitogenic effects. C1P is a direct activator of cytosolic phospholipase A2 and is involved in arachidonic acid release. CERK is a mediator of Ca2+-dependent degranulation in mast cells. In both arachidonic acid release and mast cell degranulation, the intracellular elevation of Ca2+ is a central event that acts as a regulatory mechanism of CERK activity. C1P is found in brain synaptic vesicles, and plays a role in regulating the secretion of neurotransmitters. CERK activity exists in HL-60 cells where the C1P is derived from ceramide released from sphingomyelin. The expressed kinase has specific ceramide phosphorylating activity. CERKs exist in a variety of cellular organisms, including plants, nematodes, insects, and vertebrates.

Immunogen

Full length human protein with C-terminal FLAG tag.

Positive Control

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

Formulation

Provided in 40 mM Tris-HCl pH 8.0, 110 mM NaCl, 2.2 mM KCl, 40 µg/ml Flag peptide, 3 mM DTT and 20% glycerol

Customer Storage

Product should be stored at -70°C. Aliquot to avoid freeze/thaw cycles

Target Molecular Weight

87 kDa

Database Links:

GenBankNM_022766Human

References

1. Lamour NF, Chalfant CE. ?Ceramide-1-phosphate: The missing link in eicosanoid biosynthesis and inflammation.? Mol Interv. 2005 Dec;5(6):358-67.

2.Kim JW, Inagaki Y, Mitsutake S, Maezawa N, Katsumura S, Ryu YW, Park CS, Taniguchi M, Igarashi Y. ?Suppression of mast cell degranulation by a novel ceramide kinase inhibitor, the F-12509A olefin isomer K1.? Biochim Biophys Acta. 2005 Dec 30;1738(1-3):82-90. Epub 2005 Nov 14.

3. Van Overloop H, Gijsbers S, Van Veldhoven PP. ?Further characterization of mammalian ceramide kinase: substrate delivery and (stereo)specificity, tissue distribution, and subcellular localization studies.?
J Lipid Res. 2006 Feb;47(2):268-83. Epub 2005 Nov 3.

4. Chalfant CE, Spiegel S. ?Sphingosine 1-phosphate and ceramide 1-phosphate: expanding roles in cell signaling.? J Cell Sci. 2005 Oct 15;118(Pt 20):4605-12. Review.

5. Mitsutake S, Igarashi Y. ?Calmodulin is involved in the Ca2+-dependent activation of ceramide kinase as a calcium sensor.? J Biol Chem. 2005 Dec 9;280(49):40436-41. Epub 2005 Oct 3.

6. Wijesinghe DS, Massiello A, Subramanian P, Szulc Z, Bielawska A, Chalfant CE. ?Substrate specificity of human ceramide kinase.?
J Lipid Res. 2005 Dec;46(12):2706-16. Epub 2005 Sep 18.

7.Kim TJ, Mitsutake S, Kato M, Igarashi Y. ?The leucine 10 residue in the pleckstrin homology domain of ceramide kinase is crucial for its catalytic activity.? FEBS Lett. 2005 Aug 15;579(20):4383-8.