Monoclonal antibodies generated against recombinant ATM support kinase activity

Hybridoma. 2000 Aug;19(4):317-21. doi: 10.1089/027245700429864.

Abstract

We report on the rapid generation of two monoclonal antibodies, ATM A16.35 and ATM D16.11, that bind to the kinase domain of mutated ataxia telangiectasia (ATM). These antibodies were generated against E. coli-expressed recombinant protein using the RIMMS strategy. We show that ATM A16.35 binds ATM by Western blot analysis, and ATM D16.11 forms immune complexes with native ATM in immunoprecipitations without neutralizing kinase activity.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / immunology
  • Antibody Affinity / immunology
  • Antibody Specificity / immunology
  • Ataxia Telangiectasia Mutated Proteins
  • Blotting, Western
  • Cell Cycle Proteins
  • Cell Line
  • DNA-Binding Proteins
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Immunization
  • Mice
  • Peptide Fragments
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Precipitin Tests
  • Protein-Serine-Threonine Kinases / immunology*
  • Recombinant Proteins / immunology
  • T-Lymphocytes / enzymology
  • Tumor Suppressor Proteins

Substances

  • Antibodies, Monoclonal
  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Peptide Fragments
  • Recombinant Proteins
  • Tumor Suppressor Proteins
  • Phosphatidylinositol 3-Kinases
  • ATM protein, human
  • Ataxia Telangiectasia Mutated Proteins
  • Atm protein, mouse
  • Protein-Serine-Threonine Kinases