Chip-on-beads: flow-cytometric evaluation of chromatin immunoprecipitation.
Chip-on-beads: flow-cytometric evaluation of chromatin immunoprecipitation.
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珠上芯片：染色质免疫沉淀的流式细胞术评估。
DOI：
10.1002/cyto.a.20325
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发表时间：
2006
期刊：
影响因子：
--
通讯作者：
Szabó,Gábor
中科院分区：
文献类型：
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作者：
Székvölgyi,Lóránt;Bálint,BálintL;Imre,László;Goda,Katalin;Szabó,Miklós;Nagy,László;Szabó,Gábor
BackgroundChromatin immunoprecipitation (ChIP) is a widely used technique for the detection of in vivo DNA–protein interactions underlying epigenetic regulation. The standard readout of ChIP is based on semi‐quantitative or quantitative PCR measurements; however, the development of alternative platforms with high throughput potentialities is expected to facilitate the introduction of this method into routine diagnostics.MethodsWe have established a flow‐cytometry‐based alternative for the evaluation of ChIP data. The method is based on the capture of the products of a conventional PCR run to low cycle numbers, on microbeads.ResultsIn vivo histone H4 acetylation and H3 lysine 4 methylation was detected at the promoter of the tissue transglutaminase type 2 gene. These results were confirmed by QPCR measurements. The levels of modifications decreased significantly upon apoptosis and were accompanied by the down‐regulation of TGM2 mRNA expression.ConclusionsThis method that we named ChIP‐on‐beads, a combination of flow cytometry and conventional PCR, is a reliable and efficient alternative in the quantitative analysis of ChIP results, especially promising when high throughput monitoring of epigenetic markers of diagnostic importance is required. The method is simple enough to be easily implemented in a routine flow‐cytometric laboratory. © 2006 International Society for Analytical Cytology
