title: Tyramide signal amplification (TSA) systems for the enhancement of ISH signals in cytogenetics.
abstract: This unit provides a series of techniques that dramatically improve the sensitivity of direct and indirect in situ detection. TSA is a peroxidase-based signal amplification system which is compatible with all in situ hybridization as well as immunocytochemical detection systems. The assay is performed on a glass slide and combines the use of fluorescent probes in either direct or indirect format using either an enzyme-labeled streptavidin or antibody. Protocols are provided for brightfield and fluorescent detection of single DNA targets, DNA or RNA in cultured cells, multitarget detection for DNA/RNA and DNA and RNA. In addition a protocol is included for the TSA plus system, a DNP-based detection system using horseradish peroxidase and alkaline phosphatase.
authors.affiliation: NEN Life Science Products, Boston, Massachusetts, USA.
authors.forename: M N
authors.initials: MN
authors.lastname: Bobrow
authors.forename: P T
authors.initials: PT
authors.lastname: Moen
mesh_terms: D000818:Animals; D002384:Catalysis; D003582:Cytogenetics; D004247:DNA / Q000032:analysis; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D017403:In Situ Hybridization / Q000379:methods*; D010544:Peroxidases / Q000737:chemistry; D012313:RNA / Q000032:analysis; D019809:Streptavidin / Q000737:chemistry; D014439:Tyramine / Q000032:analysis*
chemical_list: D005456:Fluorescent Dyes; D012313:RNA; D004247:DNA; D019809:Streptavidin; D010544:Peroxidases; D014439:Tyramine