MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT CCR4 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length human CCR4 cDNA (GenBank accession number NM_005508.2) 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: CCR4 (C-C chemokine receptor type 4) is a high affinity receptor for the C-C type chemokines CCL17/TARC and CCL22/MDC. CCR4 receptors express in the thymus, peripheral blood leukocytes and the spleen. CCR4 functions as a chemoattractant homing receptor on circulating memory lymphocytes and as a coreceptor for some primary HIV-2 isolates. In the CNS, CCR4 mediates hippocampal-neuron survival. CCR4 is one of the type 2 T helper cell (Th2) markers. Th2 cells are recruited to the asthmatic lung in response to allergen challenge and CCR4 has been therefore considered as a therapeutic target of lung inflammation, including eosinophilic inflammation of the lung and asthma. In addition, CCR4 has been reported as a potential independent and significant prognostic factor for adult T- cell leukemia/lymphoma (ATLL).
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.
Power and Meyer (1995) Molecular cloning and functional expression of a novel CC chemokine receptor cDNA from a human basophilic cell line. J Biol Chem 270:19495- 19500.
Imai and Baba (1997) The T cell-directed CC chemokine TARC is a highly specific biological ligand for CC chemokine receptor 4. J Biol Chem 272:15036-15042.