Oestradiol, a new hapten for detecting nucleic acid sequences by FISH.
Oestradiol has been conjugated to allylamine-dUTP with an 11-atom spacer to allow enzymatic incorporation of the label into DNA sequences. In a comparative DNA and mRNA FISH study we have used DNA probes that were either labelled with digoxigenin, biotin or oestradiol. Results show that oestradiol-labelled probes can detect DNA and RNA sequences in FISH equally well as digoxigenin- and biotin-labelled probes. Further, no crossreactivity between the various hapten-specific antibodies and the three haptens were observed. Binding of the rabbit anti-oestradiol antibody to endogenous oestrogen in various tissues was not observed under the conditions tested. In view of the increasing demands for multi-colour DNA and mRNA FISH applications, oestradiol is a welcome addition to the collection of haptens employed in FISH.
van de Corput
M P
MP
Department of Molecular Cell Biology, Leiden University, The Netherlands.
Dirks
R W
RW
Wiegant
W W
WW
Wiegant
J
J
Mühlegger
K
K
Raap
A K
AK
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics; D002460:Cell Line; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D004076:Digoxigenin / Q000737:chemistry; D004958:Estradiol / Q000031:analogs & derivatives*; D005240:Feasibility Studies; D006241:Haptens; D006367:HeLa Cells; D006801:Humans; D017874:Immediate-Early Proteins / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D015394:Molecular Structure; D012313:RNA / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis; D011817:Rabbits; D051381:Rats
D000956:Antigens, Viral; D015342:DNA Probes; D006241:Haptens; D017874:Immediate-Early Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; C044582:immediate-early proteins, cytomegalovirus; D004958:Estradiol; D012313:RNA; D004247:DNA; D004076:Digoxigenin