title: A method for simultaneous detection of multiple mRNAs using digoxigenin and radioisotopic cRNA probes.
abstract: We present a simple, reliable method for simultaneous detection of two distinct mRNAs within the same tissue sample by double in situ hybridization histochemistry. Sections are hybridized with a cocktail of radiolabeled and digoxigenin-labeled cRNA probes. The digoxigenin-labeled probe is detected with an alkaline phosphatase (AP)-dependent chromogen reaction and then the radiolabeled probe is detected by conventional autoradiography. The sensitivities of the two detection methods are comparable and demonstrate the feasibility of using double in situ hybridization histochemistry to investigate the co-localization and co-regulation of mRNAs that are expressed at relatively low levels.
authors.affiliation: Geriatric Research, Education, and Clinical Center, Veterans Affairs Medical Center, Seattle, Washington.
authors.forename: M A
authors.initials: MA
authors.lastname: Miller
authors.forename: P E
authors.initials: PE
authors.lastname: Kolb
authors.forename: M A
authors.initials: MA
authors.lastname: Raskind
mesh_terms: D000818:Animals; D001345:Autoradiography; D004076:Digoxigenin; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D018075:RNA, Complementary; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics
chemical_list: D018075:RNA, Complementary; D012333:RNA, Messenger; D004076:Digoxigenin