Improved catalyzed reporter deposition, iCARD.
Bioconjug Chem
; 25(6): 1036-42, 2014 Jun 18.
Article
en En
| MEDLINE
| ID: mdl-24852797
ABSTRACT
Novel reporters have been synthesized with extended hydrophilic linkers that in combination with polymerizing cross-linkers result in very efficient reporter deposition. By utilizing antibodies to stain HER2 proteins in a cell line model it is demonstrated that the method is highly specific and sensitive with virtually no background. The detection of HER2 proteins in tissue was used to visualize individual antigens as small dots visible in a microscope. Image analysis-assisted counting of fluorescent or colored dots allowed assessment of relative protein levels in tissue. Taken together, we have developed novel reporters that improve the CARD method allowing highly sensitive in situ detection of proteins in tissue. Our findings suggest that in situ protein quantification in biological samples can be performed by object recognition and enumeration of dots, rather than intensity-based fluorescent or colorimetric assays.
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Bencidinas
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Receptor ErbB-2
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Reactivos de Enlaces Cruzados
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Peroxidasa de Rábano Silvestre
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Anticuerpos
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
Bioconjug Chem
Asunto de la revista:
BIOQUIMICA
Año:
2014
Tipo del documento:
Article