MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT CCR1 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human CCR1 cDNA (GenBank Accession Number NM_001295) 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: CCR1 is a CC chemokine receptor with high affinity for RANTES. It is demonstrated that targeted disruption of the CCR1 receptor is associated with protection from pulmonary inflammation secondary to acute pancreatitis in the mouse. The protection from lung injury is associated with decreased levels of TNF- alpha in a temporal sequence indicating that the activation of the CCR1 receptor is an early event in the systemic inflammatory response syndrome.
Application: Functional assays
Figure 1. Dose-dependent stimulation of calcium flux upon treatment with ligand, measured with MultiscreenTM Calcium 1.0 No Wash Assay Kit (Multispan MSCA01). Figure 2. Dose-dependent inhibition of forskolin-stimulated intracellular cAMP level upon treatment with ligand, measured with MultiscreenTM TR-FRET cAMP 1.0 No Wash Assay Kit (Multispan MSCM01). 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.
Wieser et al. (2005) Expression and regulation of CCR1 in peritoneal macrophages from women with and without endometriosis. Fertil Steril 83:1878-1881.
Neote et al. (1993) Molecular cloning, functional expression, and signaling characteristics of a C-C chemokine receptor. Cell 72:415-425.