Open AccessUniversal Non‐Antibody Detection of Protein Phosphorylation Using pIMAGO
Author(s) -
Iliuk Anton B.,
Tao W. Andy
Publication year - 2015
Publication title -
current protocols in chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.503
H-Index - 14
ISSN - 2160-4762
DOI - 10.1002/9780470559277.ch140208
Subject(s) - phosphorylation , reagent , western blot , protein phosphorylation , chemistry , protein detection , antibody , microbiology and biotechnology , biochemistry , biology , nanotechnology , immunology , protein kinase a , materials science , gene
This article describes methods for a new, non‐antibody phosphorylation detection reagent, termed pIMAGO (phospho‐imaging). This novel reagent takes advantage not only of the unique properties of the soluble nanoparticles, but also of the multiple functionalities of the molecule, allowing for highly selective, sensitive, and quantitative assessment of protein phosphorylation without using radioactive isotopes or phospho‐specific antibodies. The methods allow for multiplexed detection of phosphorylation and total protein amount simultaneously. The straightforward and routine detection and quantitation of general phosphorylation on any site of any protein can be performed in western blot and ELISA formats. © 2015 by John Wiley & Sons, Inc.