Characterization of a rabbit polyclonal antibody against threonine-AMPylation

Yi Heng Hao, Trinette Chuang, Haydn L. Ball, Phi Luong, Yan Li, Ruben D. Flores-Saaib, Kim Orth

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

An antibody against the posttranslational modification AMPylation was produced using a peptide corresponding to human Rac1 switch I region with AMPylated threonine-35 residue as an antigen. The resulting rabbit antiserum was tested for its abilities to recognize AMPylated proteins by western blot and immunoprecipitation. The antiserum is highly specific for threonine-AMPylated proteins and weakly recognizes tyrosine-AMPylated proteins. Depletion of serum with modified protein abolished its activity against tyrosine-AMPylated proteins. The antiserum also recognized native proteins with modification in an immunoprecipitation experiment. Interactions of the antiserum could be inhibited by competition with AMP but not with GMP or UMP. This antiserum had potential utility for the identification of unknown AMPylated proteins.

Original languageEnglish (US)
Pages (from-to)251-254
Number of pages4
JournalJournal of Biotechnology
Volume151
Issue number3
DOIs
StatePublished - Feb 10 2011

Keywords

  • AMP
  • AMPylation
  • Antibody
  • Immunoprecipitation
  • Western blot

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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