A nonradioactive method for detecting DNA-binding activity of nuclear transcription factors / 华中科技大学学报(医学)(英德文版)
Journal of Huazhong University of Science and Technology (Medical Sciences)
;
(6): 227-9, 2003.
Artículo
en Inglés
| WPRIM
| ID: wpr-635460
ABSTRACT
To determine the feasibility of a nonradioactive electrophoresis mobility shift assay for detecting nuclear transcription factor, double-stranded oligonucleotides encoding the consensus target sequence of NF-kappa B were labelled with DIG by terminal transferase. After nuclear protein stimulated with phorbol 12-myristate 13-acetate (PMA) or PMA and pyrrolidine dithiocarbamate (PDTC) electrophoresed on 8% nondenaturing poliacrylamide gel together with oligeonucleotide probe, they were electro-blotted nylon membrane positively charged. Anti-DIG-AP antibody catalyzed chemiluminescent substrate CSPD to image on X-film. The results showed that nuclear proteins binded specifically to the NF-kappa B consensus sequence in the EMSA by chemiluminescent technique method and the activity of NF-kappa B in PMA group was more than that in PMA + PDTC group. It is suggested that detection of NF-kappa B by EMSA with chemiluminescent technique is feasible and simple, which can be performed in ordinary laboratories.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Transcripción Genética
/
Activación Transcripcional
/
Transactivadores
/
FN-kappa B
/
Ratas Sprague-Dawley
/
Ensayo de Cambio de Movilidad Electroforética
/
Proteínas de Unión al ADN
/
Mediciones Luminiscentes
Idioma:
Inglés
Revista:
Journal of Huazhong University of Science and Technology (Medical Sciences)
Año:
2003
Tipo del documento:
Artículo
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