An immunocytochemical method for the detection of fluorochrome-labelled DNA probes hybridized in situ with cellular RNA.
Various detection systems for in situ hybridization of nucleic acids are currently used. We report here an immunocytochemical detection system which is based on the detection of FITC-labelled DNA/mRNA hybrids and takes advantage of FITC molecules attached covalently to the DNA probes prior to hybridization. In situ hybridization on cytocentrifuge spots is followed by the application of an anti-fluorescein antibody thus permitting detection of mRNA/DNA-FITC hybrids. The anti-FITC antibody reaction is demonstrated by an indirect immunocytochemical peroxidase-staining method. The T lymphoblast cell line Jurkat and the cDNA for the TCR-beta chain were chosen to establish the technique.
Serke
S
S
Abteilung Innere Medizin, Klinikum R. Virchow der Freien Universität Berlin, F.R.G.
Pachmann
K
K
D002460:Cell Line; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins / Q000276:immunology*; D006801:Humans; D007124:Immunoenzyme Techniques; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis*; D011948:Receptors, Antigen, T-Cell / Q000032:analysis; D016693:Receptors, Antigen, T-Cell, alpha-beta; D013861:Thiocyanates / Q000276:immunology*
D005452:Fluoresceins; D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D016693:Receptors, Antigen, T-Cell, alpha-beta; D013861:Thiocyanates; D016650:Fluorescein-5-isothiocyanate