Time Resolved-Fluorescence Resonance Energy Transfer platform for quantitative nucleosome binding and footprinting.

ABSTRACT: Quantitative analysis of chromatin protein-nucleosome interactions is essential to understand regulation of genome-templated processes. However, current methods to measure nucleosome interactions are limited by low throughput, low signal-to-noise, and/or the requirement for specialized instrumentation. Here, we report a Lanthanide Chelate Excite Time-Resolved Fluorescence Resonance Energy Transfer (LANCE TR-FRET) assay to efficiently quantify chromatin protein-nucleosome interactions. The system makes use of commercially available reagents, offers robust signal-to-noise with minimal sample requirements, uses a conventional fluorescence microplate reader, and can be adapted for high-throughput workflows. We determined the nucleosome-binding affinities of several chromatin proteins and compl

SUBMITTER: Wesley NA 

PROVIDER: S-EPMC9134878 | biostudies-literature | 2022 Jun

REPOSITORIES: biostudies-literature