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Artículo

Development and optimization of a miniaturized western blot-based screening platform to identify regulators of post-translational modifications

Villafañez, FlorenciaIcon ; Gottifredi, VanesaIcon ; Soria, Ramiro GastonIcon
Fecha de publicación: 06/2019
Editorial: MDPI
Revista: High-Throughput
ISSN: 2571-5135
Idioma: Inglés
Tipo de recurso: Artículo publicado
Clasificación temática:
Bioquímica y Biología Molecular

Resumen

Post-translational modifications (PTMs) are fundamental traits of protein functionality and their study has been addressed using several approaches over the past years. However, screening methods developed to detect regulators of PTMs imply many challenges and are usually based on expensive techniques. Herein, we described the development and optimization of a western blot-based platform for identification of regulators of a specific PTM—mono-ubiquitylation of proliferating cell nuclear antigen (PCNA). This cell-based method does not require specific equipment, apart from the basic western blot (WB) devices and minor accessories, which are accessible for most research labs. The modifications introduced to the classical WB protocol allow the performance of PTM analysis from a single well of a 96-well plate with minimal sample manipulation and low intra- and inter-plate variability, making this method ideal to screen arrayed compound libraries in a 96-well format. As such, our experimental pipeline provides the proof of concept to design small screenings of PTM regulators by improving the quantitative accuracy and throughput capacity of classical western blots.
Palabras clave: POST-TRANSLATIONAL MODIFICATION , SCREENING , WESTERN BLOT
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info:eu-repo/semantics/openAccess Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)
Identificadores
URI: http://hdl.handle.net/11336/96521
URL: https://www.mdpi.com/2571-5135/8/2/15
DOI: http://dx.doi.org/10.3390/ht8020015
Colecciones
Articulos(CIBICI)
Articulos de CENTRO DE INV.EN BIOQUI.CLINICA E INMUNOLOGIA
Articulos(IIBBA)
Articulos de INST.DE INVEST.BIOQUIMICAS DE BS.AS(I)
Citación
Villafañez, Florencia; Gottifredi, Vanesa; Soria, Ramiro Gaston; Development and optimization of a miniaturized western blot-based screening platform to identify regulators of post-translational modifications; MDPI; High-Throughput; 8; 2; 6-2019; 1-11
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