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Open Access Gold as soon as Publ. Version/Full Text is submitted to ZB.
A multifunctional human Argonaute2-specific monoclonal antibody.
RNA 14, 1244-1253 (2008)
Small regulatory RNAs including small interfering RNAs (siRNAs), microRNAs (miRNAs), or Piwi interacting RNAs (piRNAs) guide regulation of gene expression in many different organisms. The Argonaute (Ago) protein family constitutes the cellular binding partners of such small RNAs and regulates gene expression on the levels of transcription, mRNA stability, or translation. Due to the lack of highly specific and potent monoclonal antibodies directed against the different Ago proteins, biochemical analyses such as Ago complex purification and characterization rely on overexpression of tagged Ago proteins. Here, we report the generation and functional characterization of a highly specific monoclonal anti-Ago2 antibody termed anti-Ago2(11A9). We show that anti-Ago2(11A9) is specific for human Ago2 and detects Ago2 in Western blots as well as in immunoprecipitation experiments. We further demonstrate that Ago2 can be efficiently eluted from our antibody by a competing peptide. Finally, we show that anti-Ago2(11A9) recognizes Ago2 in immunofluorescence experiments, and we find that Ago2 not only localizes to cytoplasmic processing bodies (P-bodies) and the diffuse cytoplasm but also to the nucleus. With the anti-Ago2(11A9) antibody we have generated a potent tool that is useful for many biochemical or cell biological applications.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Argonaute proteins; microRNAs; RISC; RNAi; gene silencing
Language
english
Publication Year
2008
HGF-reported in Year
2008
ISSN (print) / ISBN
1355-8382
e-ISSN
1469-9001
Journal
RNA
Quellenangaben
Volume: 14,
Issue: 6,
Pages: 1244-1253
Publisher
Cold Spring Harbor Laboratory Press
Reviewing status
Peer reviewed
Institute(s)
Institute of Molecular Immunology (IMI)
PSP Element(s)
G-501700-003
Scopus ID
44149126227
Erfassungsdatum
2008-12-12