Generation and Characterization of Novel Human IRAS Monoclonal Antibodies

Abstract EN

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.