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A proteomics approach to the study of the molecular consequence of IgE-mediated cell signalling in RBL-2H3.1 cells and 2D reference map preparation for the RBL-2H3.1 cell line

Authors

  • Esmaeil Sadroddiny,

    Corresponding author
    1. Department of Medical Biotechnology, School of Advanced Medical Technologies, Tehran University of Medical Sciences, Tehran, Iran
    2. Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, U.K.
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  • Arthur J G Moir,

    1. Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, U.K.
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  • Birgit A Helm

    1. Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield, U.K.
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To whom correspondence should be addressed (email sadroddiny@tums.ac.ir).

Abstract

A highly reproducible 2D (two-dimensional) map for the proteome and a pattern of protein phosphorylation of high secretory variant of RBL-2H3 cells (RBL-2H3.1) (a model cell in allergy studies) in resting and treated cells with IgE or IgE+Ag are presented. Major molecular changes were seen in the proteome of 3 h-activated cells with IgE+Ag, especially for proteins of ∼17 kDa compared with the control. We have identified 13 proteins on 11 corresponding spots as up-regulated proteins in response to IgE+Ag activation. Also, protein identification on 55 spots with MALDI—TOF (matrix-assisted laser-desorption ionization—time-of-flight) and ESI-MS (electrospray ionization mass spectrometry) resulted in a reliable 2D reference map and an opportunity for the subsequent use of a 1 min-activated cell map for a phosphoproteomics study.

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