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Titolo:
Dual fluorescent in situ hybridization and immunohistochemical detection with tyramide signal amplification
Autore:
Zaidi, AU; Enomoto, H; Milbrandt, J; Roth, KA;
Indirizzi:
Washington Univ, Sch Med, Dept Pathol, Div Neuropathol, St Louis, MO 63110USA Washington Univ St Louis MO USA 63110 Neuropathol, St Louis, MO 63110USA Washington Univ, Sch Med, Dept Pathol, Div Lab Med, St Louis, MO 63110 USAWashington Univ St Louis MO USA 63110 Div Lab Med, St Louis, MO 63110 USA
Titolo Testata:
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY
fascicolo: 10, volume: 48, anno: 2000,
pagine: 1369 - 1375
SICI:
0022-1554(200010)48:10<1369:DFISHA>2.0.ZU;2-N
Fonte:
ISI
Lingua:
ENG
Soggetto:
CATALYZED REPORTER DEPOSITION; INSITU HYBRIDIZATION; NEUROTROPHIC FACTOR; GDNF; SENSITIVITY; NEURTURIN; IMMUNOCYTOCHEMISTRY; RECEPTOR; NEURONS; FISH;
Keywords:
fluorescent in situ hybridization; tyramide signal amplification; immunohistochemistry; double labeling;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
39
Recensione:
Indirizzi per estratti:
Indirizzo: Roth, KA Washington Univ, Sch Med, Dept Pathol, Div Neuropathol, 660 S Euclid Ave,Box 8118, St Louis, MO 63110 USA Washington Univ 660 S Euclid Ave,Box 8118 St Louis MO USA 63110 A
Citazione:
A.U. Zaidi et al., "Dual fluorescent in situ hybridization and immunohistochemical detection with tyramide signal amplification", J HIST CYTO, 48(10), 2000, pp. 1369-1375

Abstract

To understand the biological relationships among various molecules, it is necessary to define the cellular expression patterns of multiple genes and gene products. Relatively simple methods for performing multi-label immunohistochemical detection are available. However, there is a paucity of techniques for dual immunohistochemical (IHC) and mRNA in situ hybridization (ISH) detection. The recent development of improved non-radioactive detection systems and simplified ISH protocols has prompted us to develop a tyramide signal amplification method for sequential multi-label fluorescent ISH and IHC detection in either frozen or paraffin-embedded tissue sections. We usedthis method to examine the relationship between glial cell line-derived neurotrophic factor receptor alpha 2 (GFR alpha 2) mRNA expression and IHC localization of its co-receptor Ret in the trigeminal ganglion of postnatal Day 0 mice. We found that approximately 70% of Ret-immunoreactive neurons possessed GFR alpha 2 mRNA and virtually all GFR alpha 2-expressing neurons contained Ret-immunoreactive protein. Finally, we used paraformaldehyde-fixed, paraffin-embedded sections and a monoclonal antibody against neuron-specific nuclear antigen (NeuN) to demonstrate the neuronal specificity of GFR alpha 2 mRNA expression in adult mouse brain. This multi-labeling techniqueshould be applicable to a wide variety of tissues, antibodies, and probes,providing a relatively rapid and simple means to compare mRNA and protein localization.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 12/07/20 alle ore 06:43:58