MULTISCREEN™ STABLE CELL LINES
MOUSE RECOMBINANT EBI2 RECEPTOR
1 vial (2 x 106) frozen cells
Sigma Freezing Medium (C-6164)
Expression vector containing full-length mouse EBI2 cDNA (GenBank Accession Number NM_183031.2) with FLAG tag sequence at N-terminus
Liquid nitrogen upon receiving
Propagation Medium: DMEM-F12, 10% FBS, 10 μg/mL puromycin, 250 μg/mL hygromycin
Background: Epstein-Barr virus-induced gene 2 (EBI2, also known as GPR183) is a GPCR required for humoral immune responses and polymorphisms have been associated with inflammatory autoimmune diseases. It is expressed in B-lymphocyte cell lines as well as lymphoid tissues but not T-lymphocyte cell lines or peripheral blood T-lymphocytes. 7α,25-dihydroxycholesterol (7α,25-OHC) and other oxysterols act as chemoattractants for immune cells expressing EBI2 by directing cell migration. Mice deficient in cholesterol 25- hydroxylase (CH25H, required for generation of 7α,25-OHC) fail to generate EBI2 biological activity in vivo and shows that the EBI2-oxysterol signaling pathway plays an important role in the adaptive immune response.
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. Receptor expression on cell surface measured by flow cytometry (FACS) using an anti-FLAG antibody. Thin line: parental cells; thick line: receptor-expressing cells.
Hannedouche et al. (2011) Oxysterols direct immune cell migration via EBI2. Nature 475:524-527
Birkenbach et al. (1993) Epstein-Barr virus-induced genes: first lymphocyte-specific G Protein- coupled peptide receptors. J Virol. 67(4):2209-2220.