Related Experiment Video
Updated: May 17, 2026
11:11
Flow Cytometric Analysis of Bimolecular Fluorescence Complementation: A High Throughput Quantitative Method to Study Protein-protein Interaction
Published on: August 15, 2013
A multi-parametric flow cytometric assay to analyze DNA-protein interactions
Mandana Arbab1, Shaun Mahony, Hyunjii Cho
1Division of Genetics, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115, USA.
Nucleic Acids Research
|November 13, 2012
Summary
We developed Magnetic Protein Immobilization on Enhancer DNA (MagPIE), a new assay to study DNA-protein interactions. MagPIE rapidly measures transcription factor-DNA binding and epigenetic modifications in vitro.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- Gene regulation by DNA-protein interactions is crucial for cellular function.
- Current methods to study these interactions are limited.
- Understanding transcription factor (TF)-DNA binding is essential for deciphering gene regulation.
Purpose of the Study:
- To develop a novel, rapid, and multi-parametric assay for probing DNA-protein interactions.
- To enable simultaneous analysis of TF-DNA binding, chromatin structure, and epigenetic modifications.
- To overcome limitations of existing techniques for studying gene regulation.
Main Methods:
- Magnetic Protein Immobilization on Enhancer DNA (MagPIE) assay.
- Utilizes magnetic beads bound to synthesized DNA.
- Employs flow cytometric immunofluorescence for quantitative analysis of TF-DNA binding and epigenetic markers.
Main Results:
- MagPIE allows for sequence-specific binding of TFs, histones, and methylation.
- Accurate comparative measurement of TF-DNA affinity is achievable.
- Simultaneous analysis of sequence-specific TF-DNA interaction and chromatin modification is enabled.
Conclusions:
- MagPIE is a simple, rapid, and robust method for analyzing complex epigenetic interactions in vitro.
- This assay facilitates a deeper understanding of gene regulation.
- MagPIE offers a powerful tool for studying DNA-protein interactions and their role in cellular processes.
