Development, characterisation and epitope mapping of novel monoclonal antibodies for DJ-1 (PARK7) protein

Neurosci Lett. 2005 Aug 5;383(3):225-30. doi: 10.1016/j.neulet.2005.04.024. Epub 2005 Apr 25.

Abstract

Mutations in DJ-1 (PARK7) were recently identified as the cause for an autosomal recessive early onset form of familial Parkinson's disease, however, the function of the protein in the brain is yet to be elucidated. Here we report on the development, characterisation and epitope mapping, of two novel monoclonal antibodies to DJ-1. One of them (DJ-1 "clone16") has its epitope between amino acids 56-78 of the human DJ-1 protein and has very similar properties to a commercially available DJ-1 antibody clone 3E8. The second antibody recognised both the rat and human DJ-1 (DJ-1 "clone 48") and its epitope is between amino acids 26-56. We have used immunohistochemistry with these two antibodies to compare the distribution of DJ-1 in human and rat brain tissue. Both antibodies gave similar patterns of labelling in human brain with marked astrocytic expression. Neuronal labelling was weak or absent and the antibodies did not label Lewy bodies or Lewy neurites. In the rat brain, DJ-1 was ubiquitously expressed in neurones but exhibited low expression in astrocytes. These antibodies could be exploited as important tools in dissecting out DJ-1 expression in different species and examination of the role of DJ-1 in Parkinson's disease.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aged
  • Aged, 80 and over
  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal / metabolism*
  • Antibody Specificity / physiology*
  • Antigen-Antibody Reactions / physiology
  • Blotting, Western / methods
  • Brain / metabolism
  • Epitope Mapping / methods*
  • Female
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Immunohistochemistry / methods
  • Intracellular Signaling Peptides and Proteins
  • Male
  • Mice
  • Oncogene Proteins / immunology*
  • Parkinson Disease / immunology
  • Parkinson Disease / metabolism
  • Postmortem Changes
  • Protein Deglycase DJ-1
  • Rats

Substances

  • Antibodies, Monoclonal
  • Intracellular Signaling Peptides and Proteins
  • Oncogene Proteins
  • Green Fluorescent Proteins
  • PARK7 protein, human
  • Protein Deglycase DJ-1