Fluorescent Human Delta Opioid Receptor (DOR) Internalization Assay Cell Line

Applications: Cell Screening
Categories: Cells

A novel U2OS/tGFP-DOR cell line has been developed through stable transfection to modulate the internalization of the Human Delta Opioid Receptor (DOR) complex in living cells and to assay for compounds inducing tGFP-DOR distribution inside the cells.

Fig 1:

Our expression plasmid containing the codingsequence of human Delta opioid receptortagged in the N-terminal with tGFP protein.Our plasmid was transfected in U2OS cells.Resistant clones were obtained by limitdilution, and receptor gene expression wastested by RT-PCR.

Fig 2: Internalization of DOR stimulated withDeltorphin II. Concentrations from 0 to 10 ?M were tested for1h. Activation and internalization processes were detected andanalyzed using BD Pathway 855 High-Content Bioimagerfrom BD Biosciences.

Fig 3:Concentration response curve for DeltorphinII in Delta opioid receptor cell line. Cells weretreated with 12 log dilution series (n=5). The Ec50 for theDeltorphin II was approx 3.6x10-10M after a treatment of 1 hwith the agonist. Cells were fixed and the nuclei werestained with DAPI. Percent Activity was calculated relative topositive (10 uM). The internalization assay was validatedwith an average Z prime of 0.72+/- 0.02 for High ContentScreening.