PubMed 23909421

Referenced in Channelpedia wiki pages of: none

Automatically associated channels: Kv10.1

Title: A novel monoclonal antibody against the C-terminal region of aquaporin-4.

Authors: Julia Ramadhanti, Ping Huang, Osamu Kusano-Arai, Hiroko Iwanari, Toshiko Sakihama, Tasturo Misu, Kazuo Fujihara, Takao Hamakubo, Masato Yasui, Yoichiro Abe

Journal, date & volume: Monoclon Antib Immunodiagn Immunother, 2013 Aug , 32, 270-6

PubMed link:

Aquaporin-4 (AQP4), the most abundant water channel in the brain, plays a central role in water homeostasis, neuronal activity, and migration of astrocytes in the central nervous system. Recent studies have demonstrated that AQP4 is a target of an autoantibody specifically detected in an autoimmune neurologic disease called neuromyelitis optica. Here we have generated a monoclonal antibody (MAb) against the C-terminal region of AQP4 using a baculovirus expressing mouse AQP4 as an immunogen. This antibody (clone E5206) recognized both human and mouse AQP4s in a denaturing condition and was able to precipitate AQP4 from cell lysates of CHO cells stably expressing AQP4. Western blot analysis using deletion mutants revealed that the epitope was located within a region between Asp(303) and Leu(320) in the C-terminal tail of AQP4. Although clone E5206 could not be used for immunostaining when cells or tissues were fixed with 4% paraformaldehyde or 10% formalin, it could be used when cells were fixed with 10% trichloroacetic acid and when a formalin-fixed tissue section was pretreated with antigen-retrieval reagents. This MAb can be a valuable tool for analysis of AQP4 in a variety of physiological and pathophysiological contexts, in human tissues and organs as well as in rodent models, both in vitro and in vivo.