Plant antigens cross-react with rat polyclonal antibodies against KLH-conjugated peptides.
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.
Oulehlová
Denisa
D
Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Rozvojová 263, CZ 160 00 Praha 6, Czech Republic.
Hála
Michal
M
Potocký
Martin
M
Zárský
Viktor
V
Cvrcková
Fatima
F
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000378:metabolism; D052179:Antigens, Plant / Q000276:immunology* / Q000378:metabolism; D003429:Cross Reactions; D006433:Hemocyanins / Q000276:immunology*; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology*; D051381:Rats
D000906:Antibodies; D052179:Antigens, Plant; D010455:Peptides; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin