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2D DIGE Analysis of Serum After Fractionation by ProteoMiner™ Beads

  • Cynthia Liang
  • Gek San Tan
  • Maxey C. M. ChungEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 854)

Abstract

Serum is a popular biofluid used for many protein biomarker discovery projects since the collection and processing of serum/plasma is relatively noninvasive and inexpensive. Unfortunately, the downstream analysis of serum/plasma is hampered severely by several high-abundant proteins which often interfere with the separation and detection of many of the proteins of lower abundance. Thus, a number of prefractionation methods have recently been developed with the view to reduce the dynamic range of these proteins. These include both dye- and immunoaffinity-based methods that are specifically designed to remove serum albumin. In this chapter, we describe an alternative method using ProteoMiner™ or Equalizer beads that is aimed at overcoming this problem in serum. This method uses a combinatorial library of hexapeptides bound to beads and works by binding proteins until saturation is reached. Thus, the high-abundant proteins will reach saturation quickly, while the lower-abundant proteins continue to bind. This results in a dramatic depletion of the most abundant proteins, with a concurrent concentration of the middle- to low-abundant proteins.

Key words

Serum Equalizer beads ProteoMiner™ 2D DIGE 

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Cynthia Liang
    • 1
  • Gek San Tan
    • 1
  • Maxey C. M. Chung
    • 2
    Email author
  1. 1.Department of Biological Science, Faculty of ScienceNational University of SingaporeSingaporeSingapore
  2. 2.Department of Biochemistry and Biological Sciences, Yong Loo Lin School of Medicine and Faculty of ScienceNational University of SingaporeSingaporeSingapore

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