Plant antigens cross-react with rat polyclonal antibodies against KLH-conjugated peptides

Cell Biol Int. 2009 Jan;33(1):113-8. doi: 10.1016/j.cellbi.2008.10.003. Epub 2008 Oct 22.

Abstract

Keyhole limpet hemocyanin (KLH)-conjugated peptides are routinely used to raise polyclonal antibodies for biochemical or immunolocalization studies. Rats are suitable for producing antisera against plant antigens as they often lack non-specific response towards plant materials. We attempted to obtain rat antisera against peptides derived from several plant proteins. However, most antisera recognized the same background KLH-related plant antigen (KRAP) in Arabidopsis and tobacco. We characterized KRAP with respect to size and cellular localization and examined possible antigen-specific reasons for the failure of most immunizations. We also found no reports of successful use of rat anti-KLH-peptide antibodies in plant studies. We thus believe that the rat-KLH:peptide system is poorly suited for production of antibodies, especially against plant antigens, and should be used with caution, if at all.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies / immunology*
  • Antibodies / metabolism
  • Antigens, Plant / immunology*
  • Antigens, Plant / metabolism
  • Cross Reactions
  • Hemocyanins / immunology*
  • Molecular Sequence Data
  • Peptides / immunology*
  • Rats

Substances

  • Antibodies
  • Antigens, Plant
  • Peptides
  • Hemocyanins
  • keyhole-limpet hemocyanin