Preparation of an antiserum using a fusion protein produced by a cDNA for rat aromatic L-amino acid decarboxylase

Neurosci Lett. 1993 Apr 16;153(1):88-92. doi: 10.1016/0304-3940(93)90084-x.

Abstract

Aromatic L-amino acid decarboxylase (AADC) decarboxylates L-DOPA and 5-hydroxytryptophan into dopamine and serotonin, respectively. Starting from a rat AADC cDNA clone isolated in our laboratory, we produced a beta-galactosidase-AADC fusion protein in E. coli. It was purified from inclusion bodies and injected into a rabbit. The antiserum identified AADC on a Western blot of extracts from rat organs as a unique 50 kDa band; it also strongly reacted by immunohistochemistry with dopaminergic and serotonergic neurons. This new beta-galactosidase-AADC fusion protein then constitutes a useful tool for producing AADC as an antigen free of contamination by mammalian proteins.

MeSH terms

  • Animals
  • Aromatic-L-Amino-Acid Decarboxylases / immunology*
  • Aromatic-L-Amino-Acid Decarboxylases / metabolism
  • Blotting, Western
  • Brain / metabolism
  • DNA*
  • Escherichia coli / metabolism
  • Immune Sera / immunology*
  • Immunohistochemistry
  • Rabbits
  • Rats
  • Rats, Sprague-Dawley
  • Recombinant Fusion Proteins / genetics*
  • Recombinant Fusion Proteins / immunology*
  • Recombinant Fusion Proteins / metabolism
  • Sensitivity and Specificity
  • beta-Galactosidase / immunology

Substances

  • Immune Sera
  • Recombinant Fusion Proteins
  • DNA
  • beta-Galactosidase
  • Aromatic-L-Amino-Acid Decarboxylases