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Titolo:
DOUBLE-LABELING FOR KERATIN AND CLASS-III BETA-TUBULIN WITHIN CULTURED RETINAL-PIGMENT EPITHELIAL-CELLS - COMPARISON OF CHROMOGENS TO YIELDMAXIMUM RESOLUTION OF 2 STRUCTURAL PROTEINS WITHIN THE SAME CELL
Autore:
VINORES SA; VINORES MA; CHIU C; WOERNER TM; CAMPOCHIARO PA;
Indirizzi:
JOHNS HOPKINS UNIV,SCH MED,WILMER OPTHALMOL INST,825 MAUMENEE BLDG,600 N WOLFE ST BALTIMORE MD 21287 KIRKEGAARD & PERRY LABS GAITHERSBURG MD 00000
Titolo Testata:
Journal of histotechnology
fascicolo: 1, volume: 20, anno: 1997,
pagine: 19 - 25
SICI:
0147-8885(1997)20:1<19:DFKACB>2.0.ZU;2-V
Fonte:
ISI
Lingua:
ENG
Soggetto:
UNLABELED ANTIBODY METHOD; MONOCLONAL-ANTIBODIES; EPIRETINAL MEMBRANES; 2 ANTIGENS; SIMULTANEOUS LOCALIZATION; SIMULTANEOUS EXPRESSION; ALKALINE-PHOSPHATASE; MULTIPLE ANTIGENS; COLLOIDAL GOLD; IMMUNOFLUORESCENCE;
Keywords:
CHROMOGENS; CLASS III BETA-TUBULIN; DOUBLE-LABELING; EPITHELIAL CELLS; IMMUNOHISTOCHEMISTRY; KERATIN; RETINAL PIGMENT EPITHELIAL CELLS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
46
Recensione:
Indirizzi per estratti:
Citazione:
S.A. Vinores et al., "DOUBLE-LABELING FOR KERATIN AND CLASS-III BETA-TUBULIN WITHIN CULTURED RETINAL-PIGMENT EPITHELIAL-CELLS - COMPARISON OF CHROMOGENS TO YIELDMAXIMUM RESOLUTION OF 2 STRUCTURAL PROTEINS WITHIN THE SAME CELL", Journal of histotechnology, 20(1), 1997, pp. 19-25

Abstract

Previous attempts at colocalization of 2 distinct antigens within thesame cell have met with various limitations. Either the double-labeling preparation was not permanent and required a fluorescent microscope(immuno-fluorescence) or 1 chromogen would totally or partially obscure the other, making resolution of both markers difficult. The presentstudy was conducted to assess different combinations of chromogens for optimal colocalization of 2 structural proteins, keratin and class III beta-tubulin, within cultured retinal pigment epithelial cells. Thepreferred combination of chromogens was HistoMark Red/HistoMark Blue. Good results were also achieved with HistoMark Red/3,3'-diaminobenzidine-HC1 (DAB) or True Blue, HistoMark Blue/DAB, 3-amino-9-ethylcarbazole (AEC) or HistoMark Orange, and HistoMark Orange/HistoMark Black or True Blue. It was unimportant which chromogen was used to localize each protein, but the order in which the chromogens were developed was important since some chromogens would wash out or diminish in clarity through subsequent incubations. The sequence of the chromogen application should be DAB first, then HistoMark Red, HistoMark Blue, AEC or HistoMark Orange, HistoMark Black, and lastly, 4-chloro-1-naphthol or TrueBlue.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 24/11/20 alle ore 14:10:36