Design of recombinant antibody microarrays for membrane protein profiling of cell lysates and tissue extracts

Proteomics. 2011 Apr;11(8):1550-4. doi: 10.1002/pmic.200900808. Epub 2011 Mar 17.

Abstract

Generating global protein expression profiles, including also membrane proteins, will be crucial for our understanding of biological processes in health and disease. In this study, we have expanded our antibody microarray technology platform and designed the first human recombinant antibody microarray for membrane proteins targeting crude cell lysates and tissue extracts. We have optimized all key technological parameters and successfully developed a setup for extracting, labeling and analyzing non-fractionated membrane proteomes under non-denaturing conditions. Finally, the platform was also extended and shown to be compatible with simultaneous profiling of both membrane proteins and water-soluble proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / genetics
  • Antibodies / immunology
  • Antigens, Surface / analysis*
  • Antigens, Surface / immunology
  • Female
  • Humans
  • Limit of Detection
  • Membrane Proteins / analysis*
  • Membrane Proteins / immunology
  • Palatine Tonsil / cytology
  • Placenta / cytology
  • Pregnancy
  • Protein Array Analysis / methods*
  • Recombinant Proteins
  • Reproducibility of Results
  • Tissue Extracts / analysis*
  • Tissue Extracts / immunology

Substances

  • Antibodies
  • Antigens, Surface
  • Membrane Proteins
  • Recombinant Proteins
  • Tissue Extracts