Difference gel electrophoresis of phosphoproteome

Patent metadata

Field	Value
Publication number	US-10060929-B2
Application number	US-201615004339-A
Country	US
Kind code	B2
Filing date	Jan 22, 2016
Priority date	Jan 23, 2015
Publication date	Aug 28, 2018
Grant date	Aug 28, 2018

Abstract

A novel imaging reagent and gel-based method for analyzing and comparing phosphoproteomes on the same gel are disclosed herein.

First claim

The invention claimed is:

1. A trifunctional imaging reagent, comprising:
a core linkage having the structure of

a photo-reactive group comprising diazirine;
a metal chelating group comprising a phosphate group, wherein the phosphate group has binding specifity to metal ions selected from the group consisting of Ti (VI), Zr (VI), Fe (III) and Ga (III); and
a fluorophore selected from the group consisting of Cy3, Cy5, infrared IR Dye 800 and infrared IR Dye 600, wherein the core linkage is covalently connected to the photo-reactive group, the metal chelating group and the fluorophore through an amide bond, wherein * represents the point of attachment of the fluorophore, ** represents the point of attachment of photo-reactive group and metal chelating group respectively, and wherein the metal chelating group is not linked to the core linkage through phosphate group, and photo-reactive group is not linked to the core linkage through diazirine group.

2. A kit comprising a phosphoprotein differentiating imaging reagent, the phosphoprotein differentiating imaging agent comprises a pair of the trifunctional imaging reagents of claim 1 , wherein both of the trifunctional imaging reagent in the pair are bound to the same metal ion from the selection and wherein the fluorophore in each of the trifunctional imaging reagent in the pair are selected from the selection such that the fluorophore are of different color.

Assignees

• Purdue Research Foundation

Inventors

• Tao Weiguo Andy
• Wang Linna

Classifications

• G01N2570/00

  Omics, e.g. proteomics, glycomics or lipidomics; Methods of analysis focusing on the entire complement of classes of biological molecules or subsets thereof, i.e. focusing on proteomes, glycomes or lipidomes · CPC title

• G01N33/683 Primary

  involving metal ions · CPC title

• G01N21/6428

  Measuring fluorescence of fluorescent products of reactions or of fluorochrome labelled reactive substances, e.g. measuring quenching effects, using measuring "optrodes" (in vivo A61B5/00; immunoassay G01N33/53) · CPC title

• G01N1/30

  Staining; Impregnating {; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis} · CPC title

• C07D231/56

  Benzopyrazoles; Hydrogenated benzopyrazoles · CPC title