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Beschreibung

Millipore's Ready-to-Assay GPCR frozen cells are designed for simple, rapid calcium assays with no requirement for intensive cell culturing. Millipore has optimized the freezing conditions to provide cells with high viability and functionality post-thaw. The user simply thaws the cells and resuspends them in media, dispenses cell suspension into assay plates and, following over night recovery, assays for calcium response. Sphingosine 1-phosphate (S1P) is a biologically active lysophospholipid that transmits signals through a family of five G-protein-coupled receptors to regulate cell proliferation, migration, cytoskeletal organization, and differentiation (Spiegel and Milstien , 2003). S1P4 receptor was reported to couple to multiple subsets of heterotrimeric G proteins (including Gi/o, and G12/13, and possibly Gs). Unlike other S1P receptors, S1P4 expression is restricted in human and mouse to lymph node, spleen, lung, and thymus (Ishii et al. 2001). This expression pattern suggests potential roles of S1P4 in the immune system. Millipore's cloned human S1P4-expressing cell line is made in the Chem-5 host, which supports high levels of recombinant S1P4 expression on the cell surface and contains high levels of the promiscuous G protein to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for agonists and antagonists at S1P4.