Detection and characterization of ubiquitylated H2B in mammalian cells.

Shema, Efrat; Oren, Moshe; Minsky, Neri

Shema, Efrat; Oren, Moshe; Minsky, Neri.

Affiliation

Shema E; Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel. efrat.shema@weizmann.ac.il

Methods ; 54(3): 326-30, 2011 Jul.

Article in En | MEDLINE | ID: mdl-21439383

ABSTRACT

ABSTRACT

Histone H2B ubiquitylation was shown to be associated with actively transcribed genes in mammalian cells and has been suggested to be involved in transcriptional regulation. Despite the limited applicability of genetic tools to analyze H2B ubiquitylation in mammals, several biochemical and immunological approaches have been successfully implemented to study this modification. Here we describe several techniques to detect ubiquitylated H2B in mammalian cells and to dissect its genomic localization.

Subject(s)

Histones/isolation & purification; Recombinant Fusion Proteins/isolation & purification; Ubiquitinated Proteins/isolation & purification; Analytic Sample Preparation Methods; Animals; Antibodies, Monoclonal/biosynthesis; Antibodies, Monoclonal/chemistry; Blotting, Western; Cell Extracts/chemistry; Cell Fusion; Cell Nucleus/chemistry; Cells, Cultured; Chromatin/chemistry; Chromatin/metabolism; Chromatin Immunoprecipitation; Female; Histones/chemistry; Histones/metabolism; Hybridomas; Mice; Mice, Inbred BALB C; Peptide Fragments/chemistry; Recombinant Fusion Proteins/chemistry; Recombinant Fusion Proteins/metabolism; Ubiquitinated Proteins/chemistry; Ubiquitinated Proteins/metabolism

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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Recombinant Fusion Proteins / Histones / Ubiquitinated Proteins Type of study: Diagnostic_studies Limits: Animals Language: En Journal: Methods Journal subject: BIOQUIMICA Year: 2011 Type: Article Affiliation country: Israel

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