Reverse-phase protein lysate microarrays for cell signaling analysis.
Nat Protoc
; 3(11): 1796-808, 2008.
Article
en En
| MEDLINE
| ID: mdl-18974738
ABSTRACT
'Reverse-phase' protein lysate microarray (RPA) assays use micro-scale, cell lysate dot blots that are printed to a substrate, followed by quantitative immunochemical protein detection, known to be particularly effective across many samples. Large-scale sample collection is a labor-intensive and time-consuming process; the information yielded from RPA assays, however, provides unique opportunities to experimentally interpret theoretical protein networks quantitatively. When specific antibodies are used, RPA can generate 1,000 times more data points using 10,000 times less sample volume than an ordinary western blot, enabling researchers to monitor quantitative proteomic responses for various time-scale and input-dose gradients simultaneously. Hence, the RPA system can be an excellent method for experimental validation of theoretical protein network models. Besides the initial screening of primary antibodies, collection of several hundreds of sample lysates from 1- to 8-h periods can be completed in approximately 10 d; subsequent RPA printing and signal detection steps require an additional 2-3 d.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Transducción de Señal
/
Análisis por Matrices de Proteínas
/
Péptidos y Proteínas de Señalización Intracelular
Idioma:
En
Revista:
Nat Protoc
Año:
2008
Tipo del documento:
Article
País de afiliación:
Estados Unidos