MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT P2Y12 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Full-length human P2RY12 cDNA (GenBank Accession Number NM_022788)
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin
Stable after two months of continuous growth
Background: P2RY12 or P2Y12 is a G-protein coupled receptor for ADP and ATP. ADT released from platelets acts in a positive feedback mechanism on P2Y12 and P2Y1 on platelets and is critical for sustained aggregation and stabilization of thrombin. P2Y12 receptor plays a central role in platelet activation and is the target of of antithrombotic drugs such as ticlopidine and clopidogrel.
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.
Hardy et al. (2004) Reciprocal cross-talk between P2Y1 and P2Y12 receptors at the level of calcium signaling in human platelets. Blood 104:1745-1752.
Cattaneo et al. (2003) Molecular bases of defective signal transduction in the platelet P2Y12 receptor of a patient with congenital bleeding. Proc Natl Acad Sci USA 100: 1978-1983.