A non-radioactive in situ hybridization method based on mercurated nucleic acid probes and sulfhydryl-hapten ligands
| Autor: | P.Van Duijn, Joop Wiegant, A. H. N. Hopman, G.I. Tesser |
|---|---|
| Rok vydání: | 1986 |
| Předmět: |
Pyrimidine
Organomercury Compounds Enzyme-Linked Immunosorbent Assay In situ hybridization Biology Hybrid Cells Ligands Cell Line chemistry.chemical_compound Nucleic acid thermodynamics Mice Nucleic Acids Genetics Methods Animals Humans Sulfhydryl Compounds Fluorescent Dyes Ligand Nucleic Acid Hybridization Combinatorial chemistry chemistry Biochemistry Nucleic acid Indicators and Reagents Hapten Nitrocellulose Haptens DNA |
| Zdroj: | Nucleic acids research. 14(16) |
| ISSN: | 0305-1048 |
| Popis: | Mercurated nucleic acid probes can be used for non-radioactive in situ hybridization. The principle of the method is based on the reaction of the mercurated pyrimidine residues of the in situ hybridized probe with the sulfhydryl group of a ligand which contains a hapten. Next, the hapten is immunocytochemically detected. Previous experiments showed that stable coupling of the sulfhydryl ligands could only be obtained when positively charged amino groups are present in the ligand. On basis of this finding, ligands were synthesized containing a sulfhydryl group, two lysyl residues and hapten groups such as trinitrophenyl, fluorescyl and biotinyl. The ligands, free or bound to mercurated nucleic acids, were immunochemically characterized in ELISAs. The method was shown to be specific and sensitive in the detection of target DNA in situ on microscopic preparations and in dot-blot hybridization reactions on nitrocellulose. |
| Databáze: | OpenAIRE |
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