Artículo
A fluorometric method for the assay of protein kinase activity
Rojas, Bruno Ezequiel
; Santin, Franco
; Ulloa, Rita Maria
; Iglesias, Alberto Alvaro
; Figueroa, Carlos Maria
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Fecha de publicación:
15/09/2018
Editorial:
Academic Press Inc Elsevier Science
Revista:
Analytical Biochemistry
ISSN:
0003-2697
e-ISSN:
1096-0309
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Protein kinases constitute one of the largest protein families in nature. Current methods to assay their activity involve the use of radioactive ATP or very expensive reagents. In this work, we developed a highly sensitive, cost-effective and straightforward protocol to measure protein kinase activity using a microplate layout. Released ADP is converted into NAD+, which is quantified by its fluorescent properties after alkaline treatment (linear range 0–10 nmol ADP). To validate our protocol, we characterized a recombinant calcium-dependent protein kinase from potato. Overall, this tool represents a critical step forward in the functional characterization of protein kinases.
Palabras clave:
ENZYME KINETICS
,
NON-RADIOACTIVE ASSAY
,
PHOSPHORYLATION
,
PROTEIN KINASE
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Identificadores
Colecciones
Articulos(IAL)
Articulos de INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Articulos de INSTITUTO DE AGROBIOTECNOLOGIA DEL LITORAL
Articulos(IBODA)
Articulos de INST.DE BOTANICA DARWINION (I)
Articulos de INST.DE BOTANICA DARWINION (I)
Articulos(INGEBI)
Articulos de INST.DE INVEST.EN ING.GENETICA Y BIOL.MOLECULAR "DR. HECTOR N TORRES"
Articulos de INST.DE INVEST.EN ING.GENETICA Y BIOL.MOLECULAR "DR. HECTOR N TORRES"
Citación
Rojas, Bruno Ezequiel; Santin, Franco; Ulloa, Rita Maria; Iglesias, Alberto Alvaro; Figueroa, Carlos Maria; A fluorometric method for the assay of protein kinase activity; Academic Press Inc Elsevier Science; Analytical Biochemistry; 557; 15-9-2018; 120-122
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