A novel monoclonal antibody against human Argonaute proteins reveals unexpected characteristics of miRNAs in human blood cells (original) (raw)

  1. Peter T. Nelson1,
  2. Mariangels De Planell-Saguer2,
  3. Stella Lamprinaki3,4,
  4. Marianthi Kiriakidou3,
  5. Paul Zhang2,
  6. Una O'Doherty2, and
  7. Zissimos Mourelatos2
  8. 1Department of Pathology, University of Kentucky, Lexington, Kentucky 40536, USA
  9. 2Department of Pathology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA
  10. 3Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA

Abstract

Argonaute (Ago) proteins bind to microRNA (miRNAs) and short interfering RNAs (siRNAs) and form the core components of effector complexes that mediate miRNA and siRNA function. Currently, there is a paucity of reliable antibodies against mammalian Ago proteins, thus precluding studies of endogenous Ago proteins from tissues. Here we report the development of 2A8, a novel anti-Ago monoclonal antibody that recognizes human and mouse Ago proteins and efficiently immunoprecipitates miRNAs. We report the characterization of 2A8 and its use to clone miRNAs from human brain and from preparations of human polymorphonuclear leukocytes (neutrophils), which revealed a prevalent miRNA with unusual features.

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