A method for global protein expression and antibody screening on high-density filters of an arrayed cDNA library

Nucleic Acids Res. 1998 Nov 1;26(21):5007-8. doi: 10.1093/nar/26.21.5007.


We have developed a technique to establish catalogues of protein products of arrayed cDNA clones identified by DNA hybridisation or sequencing. A human fetal brain cDNA library was directionally cloned in a bacterial vector that allows IPTG-inducible expression of His6-tagged fusion proteins. Using robot technology, the library was arrayed in microtitre plates and gridded onto high-density in situ filters. A monoclonal antibody recognising the N-terminal RGSH6sequence of expressed proteins (RGS.His antibody, Qiagen) detected 20% of the library as putative expression clones. Two example genes, GAPDH and HSP90alpha, were identified on high-density filters using DNA probes and antibodies against their proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal*
  • Brain Chemistry
  • Cloning, Molecular
  • DNA, Complementary / genetics*
  • Fetus
  • Gene Expression
  • Gene Library*
  • Humans
  • Molecular Sequence Data
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / immunology
  • Proteins / genetics*
  • Proteins / immunology*
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology


  • Antibodies, Monoclonal
  • DNA, Complementary
  • Nerve Tissue Proteins
  • Proteins
  • Recombinant Fusion Proteins

Associated data

  • GENBANK/AF074376