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Identifying RISC Components Using Ago2 Immunoprecipitation and Mass Spectrometry.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2018; Vol. 1720, pp. 149-159. - Publication Year :
- 2018
-
Abstract
- Complex immunoprecipitation (Co-IP) is a powerful technique for precipitating an intact protein complex out of solution and cell lysates using an antibody that specifically binds to a particular protein in a large complex of proteins. Mass spectrometry (MS) is used to identify, sequence, and quantify proteins. RNA-induced silencing complexes (RISCs), Ago2 centered protein assemblies, are essential for miRNA mediated RNA decay and gene expression regulation; however, the complete list of RISCs is unknown. Here we describe methods used to combine IP and MS to identify new components of RISCs.
- Subjects :
- Argonaute Proteins immunology
Argonaute Proteins metabolism
Humans
Magnetic Resonance Spectroscopy methods
MicroRNAs genetics
MicroRNAs metabolism
RNA Interference
RNA-Induced Silencing Complex genetics
Eukaryotic Initiation Factor-1 metabolism
Immunoprecipitation methods
Mass Spectrometry methods
RNA Stability genetics
RNA-Induced Silencing Complex metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 1720
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 29236257
- Full Text :
- https://doi.org/10.1007/978-1-4939-7540-2_11