2009 journal article

A streamlined method for rapid and sensitive chromatin immunoprecipitation

JOURNAL OF IMMUNOLOGICAL METHODS, 344(1), 58–63.

By: M. Sikes n, J. Bradshaw n, W. Ivory n, J. Lunsford n, R. McMillan n & C. Morrison n

author keywords: Chromatin immunoprecipitation; Histone; Epigenetics; cad; TEA; Thymocyte; Rag
MeSH headings : Animals; Aspartate Carbamoyltransferase / genetics; Aspartate Carbamoyltransferase / metabolism; Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) / genetics; Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) / metabolism; Cell Line; Chromatin / genetics; Chromatin / isolation & purification; Chromatin / metabolism; Chromatin Immunoprecipitation / methods; DNA-Binding Proteins / genetics; DNA-Binding Proteins / metabolism; Dihydroorotase / genetics; Dihydroorotase / metabolism; Gene Knockdown Techniques; Histones / genetics; Histones / metabolism; Mice; Mice, Knockout; RNA Polymerase II / genetics; RNA Polymerase II / metabolism; T-Lymphocytes / metabolism
TL;DR: A streamlined ChIP is reported to efficiently carry samples from chromatin to qPCR-compatible DNA in as little as 4 h and it is shown that the protocol readily quantified histone modifications in chromatin from 10(4) Rag-deficient DN thymocytes. (via Semantic Scholar)
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Source: Web Of Science
Added: August 6, 2018

We report a streamlined procedure to efficiently carry samples from chromatin to qPCR-compatible DNA in as little as 4 h. We use this streamlined ChIP to quantify histone H3 modifications at active (cad) and repressed (T early alpha) promoters in a Rag1-deficient pro-T cell line after 1–2 h IP. We further show that the protocol readily quantified histone modifications in chromatin from 104 Rag-deficient DN thymocytes. Taken together, these data outline a simple, cost-effective procedure for efficient ChIP analysis.