Fine epitope mapping of monoclonal antibodies specific to human alpha-synuclein.
The neuronal phosphoprotein alpha-synuclein has been increasingly implicated in the pathogenesis of Parkinson's disease (PD) and other neurodegenerative diseases; however, the exact function of alpha-synuclein still remains illusive. Suitable antibodies (Abs) specific for the gene of interest are indispensable for studying biological and immunological properties of the target gene. Here, we report not only the generation and characterization of monoclonal Abs, Syn-1 and Syn-17, against human alpha-synuclein, but also the epitope mapping by using recombinant synuclein family proteins and various GST fusion proteins of human alpha-synuclein domains. Syn-17 recognizes human and rodent alpha-synuclein, and its epitope is localized within residues 97-99 and 101 of alpha-synuclein. In contrast, the Syn-1 epitope is localized in residues 121 and 122 of human alpha-synuclein, and Syn-1 recognizes only human but not rodent alpha-synuclein, indicating that it can be utilized as a useful reagent for studying human alpha-synuclein transgenic mouse and zebrafish lines.
Choi
Ju-Youn
JY
Research Institute of Molecular Genetics, The Catholic University of Korea, Seoul 137-701, South Korea.
Park
Hyo-Jin
HJ
Seong
Young-Mo
YM
Choi
Eui Yul
EY
Min
Byung-Re
BR
Rhim
Hyangshuk
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D015153:Blotting, Western / Q000379:methods; D002460:Cell Line; D018604:Epitope Mapping; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D051379:Mice; D051381:Rats; D011994:Recombinant Proteins; D017386:Sequence Homology, Amino Acid; D014162:Transfection / Q000379:methods; D051844:alpha-Synuclein / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D051844:alpha-Synuclein