CyDye immunoblotting for proteomics: co-detection of specific immunoreactive and total protein profiles

Proteomics. 2006 Dec;6(24):6400-4. doi: 10.1002/pmic.200600139.

Abstract

The development of ECL-Plex CyDye-conjugated secondary antibodies allows the advancement of conventional Western blotting, opening up possibilities for highly sensitive and quantitative protein confirmation and identification. We report a novel proteomic method to simultaneously visualise the total protein profile as well as the specific immunodetection of an individual protein species by combining cyanine CyDye pre-labelled proteins and antibody immunoblotting. This technique proposes to revolutionise both 2-D immunoprobing and protein confirmation following MS analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Carbocyanines
  • Fluorescent Dyes
  • HSP27 Heat-Shock Proteins
  • Heat-Shock Proteins / chemistry
  • Heat-Shock Proteins / immunology
  • Heat-Shock Proteins / isolation & purification
  • Humans
  • Immunoblotting / methods*
  • Immunochemistry
  • Molecular Chaperones
  • Myocardium / chemistry
  • Neoplasm Proteins / chemistry
  • Neoplasm Proteins / immunology
  • Neoplasm Proteins / isolation & purification
  • Peptide Fragments / chemistry
  • Peptide Fragments / immunology
  • Peptide Fragments / isolation & purification
  • Protein Array Analysis / methods
  • Proteins / immunology
  • Proteins / isolation & purification
  • Proteomics / methods*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Carbocyanines
  • Fluorescent Dyes
  • HSP27 Heat-Shock Proteins
  • HSPB1 protein, human
  • Heat-Shock Proteins
  • Molecular Chaperones
  • Neoplasm Proteins
  • Peptide Fragments
  • Proteins
  • cyanine dye 3
  • cyanine dye 5