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Journal of Biomedicine and Biotechnology
Volume 2009 (2009), Article ID 973754, 8 pages
http://dx.doi.org/10.1155/2009/973754
Research Article

Generation and Characterization of Novel Human IRAS Monoclonal Antibodies

1Department of Pharmacology, Beijing Institute of Pharmacology and Toxicology, Beijing 100850, China
2Department of Pathophysiology, Beijing Institute of Radiation Medicine, Beijing 100850, China

Received 10 February 2009; Revised 18 May 2009; Accepted 5 June 2009

Academic Editor: Val J. Watts

Copyright © 2009 Bo Wang et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Imidazoline receptors were first proposed by Bousquet et al., when they studied antihypertensive effect of clonidine. A strong candidate for I1R, known as imidazoline receptor antisera-selected protein (IRAS), has been cloned from human hippocampus. We reported that IRAS mediated agmatine-induced inhibition of opioid dependence in morphine-dependent cells. To elucidate the functional and structure properties of I1R, we developed the newly monoclonal antibody against the N-terminal hIRAS region including the PX domain (10–120aa) through immunization of BALB/c mice with the NusA-IRAS fusion protein containing an IRAS N-terminal (10–120aa). Stable hybridoma cell lines were established and monoclonal antibodies specifically recognized full-length IRAS proteins in their native state by immunoblotting and immunoprecipitation. Monoclonal antibodies stained in a predominantly punctate cytoplasmic pattern when applied to IRAS-transfected HEK293 cells by indirect immunofluorescence assays and demonstrated excellent reactivity in flow immunocytometry. These monoclonal antibodies will provide powerful reagents for the further investigation of hIRAS protein functions.