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URL: https://dx.doi.org/10.17504/protocols.io.g8dbzs6

Authors: LI-COR Biosciences

Group: LI-COR Biosciences

Summary: QWB is often used for relative analysis of protein phosphorylation and other post- translational modifications (PTMs). This method combines two primary antibodies raised in different hosts: a phospho-specific or other PTM-specific antibody, and a pan-specific antibody that recognizes the target protein regardless of its modification state. Fluorescently-labeled secondary antibodies are used to simultaneously detect and discriminate the two signals. Phospho-specific signals are then normalized against the total level of the target protein, using that protein as its own internal loading control for maximum accuracy. This multiplex fluorescent method is used to analyze changes in target protein phosphorylation, glycosylation, acetylation, ubiquitination, and other PTMs. Note: This protocol is intended for use with near-infrared fluorescent Western blots.

External URL: https://www.licor.com/documents/5f2sg62r1mhpf72x50nk5rezz7clhink

Version: 1

Publication Date: 2017

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