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  Analyzing the protein assembly and dynamics of the human spliceosome with SILAC.

Schmidt, C., Raabe, M., Lührmann, R., & Urlaub, H. (2014). Analyzing the protein assembly and dynamics of the human spliceosome with SILAC. In B. Warscheid (Ed.), Stable isotope labeling by amino acids in cell culture (SILAC): Methods and protocols (pp. 227-244). New York, N.Y: Springer. doi:10.1007/978-1-4939-1142-4_16.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0023-CD5D-A Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0029-1A7B-B
Genre: Book Chapter

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 Creators:
Schmidt, C., Author
Raabe, M.1, Author              
Lührmann, R.2, Author              
Urlaub, H.1, Author              
Affiliations:
1Research Group of Bioanalytical Mass Spectrometry, MPI for biophysical chemistry, Max Planck Society, ou_578613              
2Department of Cellular Biochemistry, MPI for biophysical chemistry, Max Planck Society, ou_578576              

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Free keywords: Spliceosome , Metabolic labeling , Assembly timelines , Protein dynamics
 Abstract: Quantitative mass spectrometry has become an indispensable tool in proteomic studies. Numerous methods are available and can be applied to approach different issues. In most studies these issues include the quantitative comparison of different cell states, the identification of specific interaction partners or determining degrees of posttranslational modification. In this chapter we describe a SILAC-based quantification in order to analyze dynamic protein changes during the assembly of the human spliceosome on a pre-mRNA in vitro. We provide protocols for assembly of spliceosomes on pre-mRNA (including generation of pre-mRNAs and preparation of nuclear extracts), quantitative mass spectrometry (SILAC labeling, sample preparation), and data analysis to generate timelines for the dynamic protein assembly.

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Language(s): eng - English
 Dates: 2014-01-012014
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Method: Peer
 Identifiers: DOI: 10.1007/978-1-4939-1142-4_16
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Title: Stable isotope labeling by amino acids in cell culture (SILAC): Methods and protocols
Source Genre: Book
 Creator(s):
Warscheid, B., Editor
Affiliations:
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Publ. Info: New York, N.Y : Springer
Pages: - Volume / Issue: - Sequence Number: - Start / End Page: 227 - 244 Identifier: ISBN: 978-1-4939-1141-7

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Title: Methods in Molecular Biology
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Pages: - Volume / Issue: 1188 Sequence Number: - Start / End Page: - Identifier: -