An important way for RNA to implement its biological function in cells is the binding to proteins. Thus the analysis of RNA binding proteins in cells is necessary for the investigation of RNA's functions. So far, there have been various methods to identify RNA binding proteins. In this study, we describe a novel method, i.e. to immobilize an A/U-rich RNA to a solid medium amine M-270 magnetic beads by coupling them in a direct condensation reaction using the soluble carbodiimide reagent EDC, and then to isolate RNA binding proteins from cell lysate by RNA affinity chromatography using the immobilized RNA. The RNA specific binding proteins were then identified by SDS-PAGE, mass spectrometry and Western blotting, such as hnRNPC which bound specifically to the A/U-rich RNA of C/EBPβ 3'UTR. Colocalization in vivo between A/U-rich RNA and the identified protein hnRNPC was shown by fluorescence in situ hybridization using RNA molecular beacon and anti-hnRNPC antibody and confocal microscopy. Our results indicated that the direct immobilization of RNA with EDC is highly efficient, simple and easy.