Time-gated luminescence assay using nonmetal probes for determination of protein kinase activity-based disease markers.
Anal Biochem
; 422(2): 79-88, 2012 Mar 15.
Article
en En
| MEDLINE
| ID: mdl-22266208
ABSTRACT
A novel nonmetal optical probe ARC-1063 whose long-lifetime luminescence is induced by association with the target protein kinase is used for the measurement of the concentration of catalytic subunit of protein kinase A (PKAc) in complicated biological solutions. High affinity (K(D) = 10 pM toward PKAc) and unique optical properties of the probe enable its application for the measurement of picomolar concentrations of PKAc in the presence of high concentrations of other proteins. The described assay is applicable in the high-throughput format with the instrument setups designed for lanthanide-based time-gated (time-resolved) luminescence methods. The assay is used for demonstration that extracellular PKAc (ECPKA) is present in plasma samples of all healthy persons and cancer patients but great care must be taken for procedures of treatment of blood samples to avoid disruption, damage, or activation of platelets in the course of plasma (or serum) preparation and conservation.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Neoplasias de la Mama
/
Biomarcadores
/
Neoplasias Colorrectales
/
Proteínas Quinasas Dependientes de AMP Cíclico
/
Mediciones Luminiscentes
Tipo de estudio:
Observational_studies
/
Risk_factors_studies
Límite:
Female
/
Humans
País/Región como asunto:
Europa
Idioma:
En
Revista:
Anal Biochem
Año:
2012
Tipo del documento:
Article
País de afiliación:
Estonia