MULTISCREEN™ STABLE CELL LINES
MOUSE RECOMBINANT BLT2 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length mouse BLT2 cDNA (GenBank Accession Number NM_020490.2) 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: Leukotriene B4 (LTB4) is a potent lipid mediator of allergic and inflammatory reactions, as well as a modulator of immune responses. In mice, BLT2 receptor has been shown to be expressed in the small intestine and skin and functional BLT2 in primary keratinocytes, compared to the ubiquitous expression in humans. Cells expressing BLT2 exhibited LTB4-induced chemotaxis, calcium mobilization, and inhibition of adenylyl cyclase. In an autoantibody-induced inflammatory arthritis model, BLT2-knockout mice showed decreased incidence and severity of disease. Thus, BLT2 provides a novel target for anti-inflammatory therapy and promises to expand our knowledge of LTB4 function.
Application: Functional assays
Figure 1. 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 2. Dose-dependent stimulation of calcium flux upon treatment with ligand, measured with MultiscreenTM Calcium 1.0 No Wash Assay Kit (Multispan MSCA01). 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.
Iizuka et al. (2005) Characterization of a mouse second leukotriene receptor, mBLT2: BLT2-dependent ERK activation and cell migration of prim,ary mouse keratinocytes. J Biol Chem 280(26):24816-23.
Mathis et al. (2010) Nonredundant roles for leukotriene B4 receptors BLT1 and BLT2 in inflammatory arthritis. J Immunol. 185(5):3049-56.
Yokomizo et al. (2000) A second leukotriene B(4) receptor, BLT2. A new therapeutic target in inflammation and immunological disorders. J Exp Med 192:421-432.