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Double One-Dimensional Electrophoresis (D1-DE) Adapted for Immunoproteomics. | LitMetric

Double One-Dimensional Electrophoresis (D1-DE) Adapted for Immunoproteomics.

Methods Mol Biol

Allergy and Environment Team, Biochemistry Department, Armand Trousseau Children Hospital (AP-HP), Paris, France.

Published: June 2019

AI Article Synopsis

  • * This traditional method requires running at least two gels, which can be time-consuming and uses a lot of reagents for protein purification and characterization.
  • * The modified double one-dimensional electrophoresis (D1-DE) technique offers a more efficient way to convert 2-DE protein spots into larger protein bands, increasing protein abundance for further functional analyses from a single separation.

Article Abstract

The classical proteomics approach for the identification of allergen candidates consists on the separation of proteins by high-resolution two-dimensional electrophoresis (2-DE) with subsequent IgE immunoblotting and further analysis of IgE-reactive protein spots with mass spectrometry. In this approach at least two gels most be run. One gel is used for staining and the other is for immunoblotting by antibodies labeled with specific immunostains. Additional functional characterizations require either protein purification or 2-DE replicates and appear to be time- and reagent-consuming. Here we described a modified double one-dimensional electrophoresis (D1-DE) allowing the conversion of a protein spot previously visualized by 2-DE into an extended protein band. In D1-DE, the purity of the protein of interest is similar to 2-DE spots, but its abundance is many times higher than what can be found in a 2-DE single spot allowing many other functional analyses from a single D1-DE separation.

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Source
http://dx.doi.org/10.1007/978-1-4939-8814-3_9DOI Listing

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