Zymography on peroxidase-entrapped gels enables the determination of the intrinsic specific activity (ISA) of H2O2-producing oxidases in the presence of interfering enzymes including catalase (decomposing hydrogen peroxide). To reveal the searched oxidase, the zymography gel is incubated first in the developing solution containing the appropriate substrate of the oxidase and an oxidizable chromogen such as o-phenylenediamine (o-PDA) peroxidase substrate. The zymographic gel, after image analysis, can be restained by Coomassie Blue for molecular weights, for the whole electrophoretic pattern and for the protein concentration, thus allowing for the determination of oxidase ISA.
Zymographic determination of intrinsic specific activity of oxidases in the presence of interfering proteins / Le Tien, Canh; Mateescu Mircea, Alexandru; Ahmadifar, Samaneh; Marcocci, Lucia; Pietrangeli, Paola. - STAMPA. - (2018), pp. 207-221. - METHODS IN MOLECULAR BIOLOGY. [10.1007/978-1-4939-8745-0_24].
Zymographic determination of intrinsic specific activity of oxidases in the presence of interfering proteins
Marcocci Lucia;Pietrangeli Paola
2018
Abstract
Zymography on peroxidase-entrapped gels enables the determination of the intrinsic specific activity (ISA) of H2O2-producing oxidases in the presence of interfering enzymes including catalase (decomposing hydrogen peroxide). To reveal the searched oxidase, the zymography gel is incubated first in the developing solution containing the appropriate substrate of the oxidase and an oxidizable chromogen such as o-phenylenediamine (o-PDA) peroxidase substrate. The zymographic gel, after image analysis, can be restained by Coomassie Blue for molecular weights, for the whole electrophoretic pattern and for the protein concentration, thus allowing for the determination of oxidase ISA.File | Dimensione | Formato | |
---|---|---|---|
Le_Zymographic_2018.pdf
solo gestori archivio
Tipologia:
Versione editoriale (versione pubblicata con il layout dell'editore)
Licenza:
Tutti i diritti riservati (All rights reserved)
Dimensione
2.43 MB
Formato
Adobe PDF
|
2.43 MB | Adobe PDF | Contatta l'autore |
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.