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  Chromatin Immunoprecipitation (ChIP) of Heat Shock Protein 90 (Hsp90)

Yoveva, A., & Sawarkar, R. (2017). Chromatin Immunoprecipitation (ChIP) of Heat Shock Protein 90 (Hsp90). In Methods in Molecular Biology (pp. 221-231). Springer Science + Business Media.

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 Urheber:
Yoveva, Aneliya1, Autor
Sawarkar, Ritwick1, Autor           
Affiliations:
1Max Planck Institute of Immunobiology and Epigenetics, Max Planck Society, 79108 Freiburg, DE, ou_2243640              

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Schlagwörter: ChIP-Seq, Drosophila Schneider (S2) Cells, Hsp90
 Zusammenfassung: Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a widely used technique for genome-wide mapping of protein-DNA interactions and epigenetic marks in vivo. Recent studies have suggested an important role of heat shock protein 90 (Hsp90) at chromatin. This molecular chaperone assists other proteins to acquire their mature and functional conformation and helps in the assembly of many complexes. In this chapter, we provide specific details on how to perform Hsp90 ChIP-seq from Drosophila Schneider (S2) cells. Briefly, the cells are simultaneously lyzed and reversibly cross-linked to stabilize protein-DNA interactions. Chromatin is prepared from isolated nuclei and sheared by sonication. Hsp90-bound loci are immunoprecipitated and the corresponding DNA fragments are purified and sequenced. The described approach revealed that Hsp90 binds close to the transcriptional start site of around one-third of all Drosophila coding genes and characterized the role of the chaperone at chromatin.

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Sprache(n): eng - English
 Datum: 2017
 Publikationsstatus: Erschienen
 Seiten: -
 Ort, Verlag, Ausgabe: -
 Inhaltsverzeichnis: -
 Art der Begutachtung: Expertenbegutachtung
 Identifikatoren: DOI: 10.1007/978-1-4939-7477-1_17
 Art des Abschluß: -

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Titel: Methods in Molecular Biology
Genre der Quelle: Buch
 Urheber:
Affiliations:
Ort, Verlag, Ausgabe: Springer Science + Business Media
Seiten: - Band / Heft: 1709 Artikelnummer: - Start- / Endseite: 221 - 231 Identifikator: -