Technical Reference #445

445.

Real-Time Optical Monitoring Of Ligand-Mediated Internalization Of A1B-Adrenoceptor With Green Fluorescent Protein Takeo Awaji; Akira Hirasawa; Masakazu Kataoka; Hitomi Shinoura; Yasuhisa Nakayama; Tatsuo Sugawara; Shun-ichiro Izumi; and Gozoh Tsujimoto, National Children's Medical Research Center, Molecular Endocrinology, 12(445), (1998)
Link To Paper

Abstract:
The study of G protein-coupled receptor signaltransduction and behavior in living cells is technicallydifficult because of a lack of useful biologicalreagents. We show here that a fully functional a1b-adrenoceptor tagged with the green fluorescent

Keywords:
agonist-promoted a1bAR/GFP redistribution; a1-Adrenoceptors; phospholipase C; Green fluorescent protein

Materials & Methods:
aT3 cells stably expressing wild-type a1bAR and a1bAR/GFP were seeded at 1 3 105 per well of the cover glass-bottom culture dish (MatTek Corp. Ashland MA) in 2.0 ml of medium and examined using LSM-GB200 within 30 min at room temperature.

Microscopic Technique
Confocal Microscopy

Cell Type(s)
AT3