A Nanomolar-Affinity Antibody Enabling Sensitive Labeling and High-Resolution Genomic Mapping of O-GlcNAc

Here, we report the development of a pan-specific, nanomolar-affinity O-GlcNAc antibody named EPR19847 by using a degenerate O‑GlcNAcylated peptide as the immunogen. The EPR19847 antibody can be used for O‑GlcNAc detection, imaging, isolation, and chromatin immunoprecipitation.

Using this novel antibody, O-GlcNAcylated proteins were enriched and identified in mouse embryonic stem cells (mESCs). Chromatin immunoprecipitation followed by next-generation sequencing (ChIP-seq) demonstrates that O-GlcNAc is enriched around the promoters of highly expressed genes, and that its genomic distribution is positively correlated with transcription-activating marks including H3K4me3, H3K9ac, and RNA Pol II in mESCs.