title: Generation and characterization of novel human IRAS monoclonal antibodies.
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.
authors: Department of Pharmacology, Beijing Institute of Pharmacology and Toxicology, Beijing 100850, China.
authors: Bo
authors: B
authors: Wang
authors: Ying
authors: Y
authors: Liu
authors: Yajun
authors: Y
authors: Shan
authors: Zhenyu
authors: Z
authors: Yao
authors: Xiaolan
authors: X
authors: Liu
authors: Ruibin
authors: R
authors: Su
authors: Qihong
authors: Q
authors: Sun
authors: Yuwen
authors: Y
authors: Cong
authors: Jin
authors: J
authors: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology / Q000378:metabolism; D002460:Cell Line; D004926:Escherichia coli / Q000235:genetics; D005434:Flow Cytometry; D006801:Humans; D006825:Hybridomas / Q000378:metabolism; D054755:Imidazoline Receptors / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D015151:Immunoblotting; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D054755:Imidazoline Receptors; D047908:Intracellular Signaling Peptides and Proteins; C458505:NISCH protein, human; D011993:Recombinant Fusion Proteins