Differential Adipose Tissue Proteomics

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


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).


Adipose tissue Mass spectrometry Proteomics Western blotting 


  1. 1.
    Geguchadze RN, Machen L, Zourelias L, Gallo PH, Passineau MJ (2012) An AAV2/5 vector enhances safety of gene transfer to the mouse salivary gland. J Dent Res 91:382–386CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Shields KJ, Verdelis K, Passineau MJ, Faight EM, Zourelias L, Wu C et al (2016) Three-dimensional micro computed tomography analysis of the lung vasculature and differential adipose proteomics in the Sugen/hypoxia rat model of pulmonary arterial hypertension. Pulm Circ 6:586–596CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Calderón-Celis F, Encinar JR, Sanz-Medel A (2017) Standardization approaches in absolute quantitative proteomics with mass spectrometry. Mass Spec Rev 9999:1–23.
  4. 4.
    Trayhurn P (2005) Endocrine and signalling role of adipose tissue: new perspectives on fat. Acta Physiol Scand 184:285–293CrossRefPubMedGoogle Scholar
  5. 5.
    Baker AR, Silva NF, Quinn DW, Harte AL, Pagano D, Bonser RS et al (2006) Human epicardial adipose tissue expresses a pathogenic profile of adipocytokines in patients with cardiovascular disease. Cardiovasc Diabetol 5:1CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Cypess AM, Lehman S, Williams G, Tal I, Rodman D, Goldfine AB et al (2009) Identification and importance of brown adipose tissue in adult humans. N Engl J Med 360:1509–1517CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Sharp LZ, Shinoda K, Ohno H, Scheel DW, Tomoda E, Ruiz L et al (2012) Human BAT possesses molecular signatures that resemble beige/brite cells. PLoS One 7:e49452CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Peinado JR, Pardo M, de la Rosa O, Malagon MM (2012) Proteomic characterization of adipose tissue constituents, a necessary step for understanding adipose tissue complexity. Proteomics 12:607–620CrossRefPubMedGoogle Scholar
  9. 9.
    Chen CL, Chung T, CC W, Ng KF, JS Y, Tsai CH et al (2015) Comparative tissue proteomics of microdissected specimens reveals novel candidate biomarkers of bladder cancer. Mol Cell Proteomics 14:2466–2478CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Wu C, Wang Z, Zourelias L, Thakker H, Passineau MJ (2015) IL-17 sequestration via salivary gland gene therapy in a mouse model of Sjogren’s syndrome suppresses disease-associated expression of the putative autoantigen Klk1b22. Arthritis Res Ther 17:198CrossRefPubMedPubMedCentralGoogle Scholar

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

Personalised recommendations