MULTISCREEN™ STABLE CELL LINES
HUMAN RECOMBINANT BLT2 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length monkey BLT2 cDNA (GenBank Accession Number XM_001113942.2) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin
Background: Leukotriene B4 (LTB4) is a potent lipid mediator of allergic and inflammatory reactions, as well as a modulator of immune responses. Two receptors for LTB4 have been described on human neutrophils. The high-affinity human leukocyte LTB4 receptor, BLT1 mediates aggregation, chemotaxis, chemokinesis, and increased adherence to surfaces, whereas the low-affinity receptor, BLT2 (or LTB4R2) mediates degranulation and increased oxidative metabolism. BLT2 is expressed ubiquitously with the highest expression in spleen and has a broader ligand specificity for various eicosanoids. Cells expressing BLT2 exhibited LTB4-induced chemotaxis, calcium mobilization, and inhibition of adenylyl cyclase. 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.
Kamohara et al. (2000) Molecular cloning and characterization of another leukotriene B4 receptor. J Biol Chem 275:27000-27004.
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.
Yoo et al. (2004) Role of the BLT2, a leukotriene B4 receptor, in Ras transformation. Oncogene 23:9259-9268.