ChIP-on-beads: Flow-cytometric evaluation of chromatin immunoprecipitation

Lóránt Székvölgyi, Bálint L. Bálint, László Imre, K. Goda, Miklós Szabó, L. Nagy, Gábor Szabó

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: Chromatin 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. Methods: We 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. Results: In 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. Conclusions: This method that we named ChIP-onbeads, 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.

Original languageEnglish
Pages (from-to)1086-1091
Number of pages6
JournalCytometry Part A
Volume69
Issue number10
DOIs
Publication statusPublished - Oct 2006

Fingerprint

Chromatin Immunoprecipitation
Epigenomics
Polymerase Chain Reaction
Histones
Flow Cytometry
Acetylation
Microspheres
Methylation
Lysine
Down-Regulation
Apoptosis
Messenger RNA
DNA
Genes
Proteins
transglutaminase 2

Keywords

  • Chromatin immunoprecipitation
  • Flow cytometry

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

Cite this

Székvölgyi, L., Bálint, B. L., Imre, L., Goda, K., Szabó, M., Nagy, L., & Szabó, G. (2006). ChIP-on-beads: Flow-cytometric evaluation of chromatin immunoprecipitation. Cytometry Part A, 69(10), 1086-1091. https://doi.org/10.1002/cyto.a.20325

ChIP-on-beads : Flow-cytometric evaluation of chromatin immunoprecipitation. / Székvölgyi, Lóránt; Bálint, Bálint L.; Imre, László; Goda, K.; Szabó, Miklós; Nagy, L.; Szabó, Gábor.

In: Cytometry Part A, Vol. 69, No. 10, 10.2006, p. 1086-1091.

Research output: Contribution to journalArticle

Székvölgyi, L, Bálint, BL, Imre, L, Goda, K, Szabó, M, Nagy, L & Szabó, G 2006, 'ChIP-on-beads: Flow-cytometric evaluation of chromatin immunoprecipitation', Cytometry Part A, vol. 69, no. 10, pp. 1086-1091. https://doi.org/10.1002/cyto.a.20325
Székvölgyi, Lóránt ; Bálint, Bálint L. ; Imre, László ; Goda, K. ; Szabó, Miklós ; Nagy, L. ; Szabó, Gábor. / ChIP-on-beads : Flow-cytometric evaluation of chromatin immunoprecipitation. In: Cytometry Part A. 2006 ; Vol. 69, No. 10. pp. 1086-1091.
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