期刊
PROTEOMICS
卷 10, 期 16, 页码 3045-3050出版社
WILEY
DOI: 10.1002/pmic.201000162
关键词
Animal proteomics; Nucleolus; Protein phosphatase; Quantitative proteomics; Stable isotope labeling by amino acids in cell culture
资金
- NSERC [372370]
- Terry Fox Foundation [20148]
The efficient extraction of proteins from purified cellular organelles is critical for in vitro analyses, including identification of protein complex members by affinity purification-based quantitative proteomic approaches. When applied to purified nucleoli, classic nuclear protein extraction methods inefficiently and selectively release only similar to 50% of proteins. Here, we present a method that can extract up to 90% of nucleolar proteins, and apply it in a quantitative interactomic approach to identify nucleolar interaction partners for a mammalian protein phosphatase.
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