A novel monoclonal antibody against human Argonaute proteins reveals unexpected characteristics of miRNAs in human blood cells.
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.
Nelson
Peter T
PT
Department of Pathology, University of Kentucky, Lexington, KY 40536, USA. peter.nelson@uky.edu
De Planell-Saguer
Mariangels
M
Lamprinaki
Stella
S
Kiriakidou
Marianthi
M
Zhang
Paul
P
O'Doherty
Una
U
Mourelatos
Zissimos
Z
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001483:Base Sequence; D002460:Cell Line; D015852:Eukaryotic Initiation Factor-2 / Q000235:genetics*; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism*; D034622:RNA Interference; D011994:Recombinant Proteins / Q000378:metabolism
D000911:Antibodies, Monoclonal; D015852:Eukaryotic Initiation Factor-2; D035683:MicroRNAs; D011994:Recombinant Proteins