MULTISCREEN™ STABLE CELL LINES
MOUSE RECOMBINANT CCR6 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length mouse Ccr6 DNA (GenBank Accession Number BC100758) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin
Stable in culture for minimum of two months
Background: CCR6 is a receptor for macrophage inflammatory protein (MIP)-3α (CCL20). It is expressed in immature dendritic cells as well as in B lymphocytes and memory T cells. In gene-targeted mice lacking CCR6, myeloid dendritic cells are either absent or underrepresented in the subepithelial dome compared with wild-type mice. CCR6 and its ligands contribute to the normal homeostatic regulation of extrathymic T cell development in the intestine.
Application: Functional assays
Figure 1. Dose-dependent inhibition of forskolin-stimulated cAMP level upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). Figure 2. Dose-dependent stimulation of calcium flux upon treatment with ligand, measured with MultiscreenTM Calcium 1.0 No Wash Assay Kit (Multispan MSCA01). Cells were transiently transfected with Gαqi5. Figure 3. Receptor expression on cell surface measured by flow cytometry (FACS) using an anti-FLAG antibody. Thin line: parental cells; thick line: receptor-expressing cells.
Baba et al. (1997) Identification of CCR6, the specific receptor for a novel lymphocytedirected CC chemokine LARC. J Biol Chem 272:14893-14898.
Cook et al. (2000) CCR6 mediates dendritic cell localization, lymphocyte homeostasis, and immune responses in mucosal tissue. Immunity 12:495-503.
Lugering et al. (2003) Lymphoid precursors in intestinal cryptopatches express CCR6 and undergo dysregulated development in the absence of CCR6. J Immunol171:2208-2215.