TY - JOUR
T1 - 2-Acetylaminofluorene-modified probes for the indirect hybridocytochemical detection of specific nucleic acid sequences
AU - Landegent, J. E.
AU - De Wal, N. Jansen In
AU - Baan, R. A.
AU - Hoeijmakers, J. H.J.
AU - Van Der Ploeg, M.
N1 - Funding Information:
The authors are very grateful to Dr P. Tchen who pointed out the idea of using AAF-modified probes for hybridization purposes. We thank Professor P. Van Duijn and Professor P. Borst for their encouragement and helpful discussions. We also thank Dr 0. B. Zaalberg and his colleagues for immunizing the rabbits and preparing the antiserum, and Mr C. E. Kientz for providing us with an HPLC RSil column. Part of this work was supported by the Netherlands Organisation of the Advancement of Pure Research (Z.W.O.).
PY - 1984/7
Y1 - 1984/7
N2 - A new approach is presented for the indirect hybridocytochemical localization of specific nucleic acid sequences in microscopic preparations. The method is based on the application of probes modified with N-acetoxy-2-acetylaminofluorene. After hybridization, the 2-acetylaminofluorene-labelled probes are recognized by antibodies directed against modified guanosine and visualized immunocytochemically. This procedure has been optimized on two model objects: mouse satellite DNA in interphase nuclei and chromosomes, and kinetoplast DNA in Crithidia fasciculata. A first application that may be of clinical importance is given by the detection of human cytomegalovirus in infected human lung fibroblasts. Other potentials of this procedure are discussed. Its advantages are: (1) the simple, rapid and reproducible labelling procedure; (2) the high stability of both label and modified probes; (3) the feasibility of labelling both double-stranded (ds) and single-stranded (ss) probes (DNA as well as RNA); (4) the rapid and sensitive detection of hybrids.
AB - A new approach is presented for the indirect hybridocytochemical localization of specific nucleic acid sequences in microscopic preparations. The method is based on the application of probes modified with N-acetoxy-2-acetylaminofluorene. After hybridization, the 2-acetylaminofluorene-labelled probes are recognized by antibodies directed against modified guanosine and visualized immunocytochemically. This procedure has been optimized on two model objects: mouse satellite DNA in interphase nuclei and chromosomes, and kinetoplast DNA in Crithidia fasciculata. A first application that may be of clinical importance is given by the detection of human cytomegalovirus in infected human lung fibroblasts. Other potentials of this procedure are discussed. Its advantages are: (1) the simple, rapid and reproducible labelling procedure; (2) the high stability of both label and modified probes; (3) the feasibility of labelling both double-stranded (ds) and single-stranded (ss) probes (DNA as well as RNA); (4) the rapid and sensitive detection of hybrids.
UR - http://www.scopus.com/inward/record.url?scp=0021154086&partnerID=8YFLogxK
U2 - 10.1016/0014-4827(84)90448-8
DO - 10.1016/0014-4827(84)90448-8
M3 - Article
C2 - 6203769
AN - SCOPUS:0021154086
SN - 0014-4827
VL - 153
SP - 61
EP - 72
JO - Experimental Cell Research
JF - Experimental Cell Research
IS - 1
ER -