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Differential Adipose Tissue Proteomics

  • Kelly J. ShieldsEmail author
  • Changgong Wu
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1788)

Abstract

Differential proteomic analysis (comparative quantitative proteomics) is a robust quantitative technique used to detect and identify the proteome of selected tissues. The expression levels (upregulated vs. downregulated) of proteins in tissue samples that differ by experimental design or anatomic location are determined by a series of assays including (1) 2D difference gel electrophoresis (2D-DiGE), (2) protein spot picking based on a priori thresholds, (3) Mass Spectrometry, and (4) follow-up Western Blot for antibody validation (Chen et al., Mol Cell Proteomics 14:2466–2478, 2015). Differential proteomic analysis is a perfect method for analyzing a heterogeneous tissue such as adipose tissue with a composition spectrum consisting of white to brown adipocytes along with a stromal vascular fraction dependent on anatomical location and inflammation. The adipose tissue proteomic protocol outlined here was successful in identifying differentially expressed proteins both significantly upregulated and downregulated between the experimental and control groups (Shields et al., Pulm Circ 6:586–596, 2016).

Keywords

Adipose tissue Mass spectrometry Proteomics Western blotting 

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of MedicineLupus Center of Excellence – Autoimmunity Institute, Allegheny Health NetworkPittsburghUSA
  2. 2.Allegheny General Hospital320 E North Avenue, 8th FL South TowerPittsburghUSA
  3. 3.Center for Advanced Proteomics Research, New Jersey Medical School Rutgers, The State University of New JerseyNewarkUSA

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