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Chemically-blocked Antibody Microarray for Multiplexed High-throughput Profiling of Specific Protein Glycosylation in Complex Samples
Published on: May 4, 2012
A multifunctional human Argonaute2-specific monoclonal antibody
Sabine Rüdel1, Andrew Flatley, Lasse Weinmann
1Center for Integrated Protein Science Munich, Laboratory of RNA Biology, Max Planck Institute of Biochemistry, 82152 Martinsried, Germany.
Researchers developed a specific monoclonal antibody, anti-Ago2(11A9), for human Argonaute-2 (Ago2) protein. This antibody enables precise detection and analysis of Ago2 in various biochemical and cell biology applications, including its nuclear localization.
Area of Science:
- Molecular Biology
- Genetics
- Immunology
Background:
- Small regulatory RNAs (siRNAs, miRNAs, piRNAs) are crucial for gene expression regulation.
- Argonaute (Ago) proteins are key binding partners of small RNAs, influencing gene expression at multiple levels.
- A lack of specific antibodies against Ago proteins hinders biochemical analysis, often necessitating protein overexpression.
Purpose of the Study:
- To generate and functionally characterize a highly specific monoclonal antibody against human Ago2.
- To provide a reliable tool for biochemical and cell biological studies of Ago2.
Main Methods:
- Generation of a monoclonal antibody, anti-Ago2(11A9).
- Validation of antibody specificity using Western blot and immunoprecipitation.
- Demonstration of antigen elution via competing peptide.
- Immunofluorescence microscopy to determine Ago2 localization.
Main Results:
- The anti-Ago2(11A9) antibody specifically recognizes human Ago2.
- The antibody is effective in Western blot and immunoprecipitation assays.
- Ago2 can be efficiently eluted from the antibody using a competing peptide.
- Immunofluorescence revealed Ago2 localization in the cytoplasm, P-bodies, and the nucleus.
Conclusions:
- The anti-Ago2(11A9) antibody is a specific and potent tool for studying human Ago2.
- This antibody facilitates various biochemical and cell biological applications, including localization studies.
- The findings confirm Ago2's presence in both cytoplasmic and nuclear compartments.
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