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 human BLT2 cDNA (GenBank Accession Number NM_019839.1) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM, 10% FBS, 1 μg/mL puromycin
Stable after minimum of 2 months continuous growth
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 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.
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.