Artículo

Rojas, B.E.; Santin, F.; Ulloa, R.M.; Iglesias, A.A.; Figueroa, C.M. "A fluorometric method for the assay of protein kinase activity" (2018) Analytical Biochemistry. 557:120-122
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Abstract:

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. © 2018 Elsevier Inc.

Registro:

Documento: Artículo
Título:A fluorometric method for the assay of protein kinase activity
Autor:Rojas, B.E.; Santin, F.; Ulloa, R.M.; Iglesias, A.A.; Figueroa, C.M.
Filiación:Instituto de Agrobiotecnología del Litoral, UNL, CONICET, FBCB, Colectora Ruta Nacional 168 Km 0, Santa Fe, 3000, Argentina
Instituto de Investigaciones en Ingeniería Genética y Biología Molecular, UBA, CONICET, FCEN, Vuelta de Obligado 2490, Buenos Aires, 1428, Argentina
Palabras clave:Enzyme kinetics; Non-radioactive assay; Phosphorylation; Protein kinase; lactate dehydrogenase; protein kinase; pyruvate kinase; Article; enzyme activity; fluorometry; kinetic parameters; limit of detection; potato; priority journal; substrate concentration
Año:2018
Volumen:557
Página de inicio:120
Página de fin:122
DOI: http://dx.doi.org/10.1016/j.ab.2018.07.018
Título revista:Analytical Biochemistry
Título revista abreviado:Anal. Biochem.
ISSN:00032697
CODEN:ANBCA
CAS:lactate dehydrogenase, 9001-60-9; protein kinase, 9026-43-1; pyruvate kinase, 9001-59-6
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00032697_v557_n_p120_Rojas

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Citas:

---------- APA ----------
Rojas, B.E., Santin, F., Ulloa, R.M., Iglesias, A.A. & Figueroa, C.M. (2018) . A fluorometric method for the assay of protein kinase activity. Analytical Biochemistry, 557, 120-122.
http://dx.doi.org/10.1016/j.ab.2018.07.018
---------- CHICAGO ----------
Rojas, B.E., Santin, F., Ulloa, R.M., Iglesias, A.A., Figueroa, C.M. "A fluorometric method for the assay of protein kinase activity" . Analytical Biochemistry 557 (2018) : 120-122.
http://dx.doi.org/10.1016/j.ab.2018.07.018
---------- MLA ----------
Rojas, B.E., Santin, F., Ulloa, R.M., Iglesias, A.A., Figueroa, C.M. "A fluorometric method for the assay of protein kinase activity" . Analytical Biochemistry, vol. 557, 2018, pp. 120-122.
http://dx.doi.org/10.1016/j.ab.2018.07.018
---------- VANCOUVER ----------
Rojas, B.E., Santin, F., Ulloa, R.M., Iglesias, A.A., Figueroa, C.M. A fluorometric method for the assay of protein kinase activity. Anal. Biochem. 2018;557:120-122.
http://dx.doi.org/10.1016/j.ab.2018.07.018