Poster Presentation 51st Lorne Proteins Conference 2026

Quantitative Profiling of Endogenous Protein Interactions Using HiBiT Tag and IP-MS (#230)

David Boucher 1 , Virginia Kincaid 2 , Christopher Eggers 2 , Mike Rosenblatt 2 , Brock Binkowski 2 , Marjeta Urh 2
  1. Promega Corporation, Alexandria, VIC, Australia
  2. Promega Corporation, Madison, USA

HiBiT is an 11-amino-acid peptide tag that enables rapid and sensitive quantification in vitro or in live cells. HiBiT is detected by bioluminescent enzyme complementation in simple, add-and-read assays. The higher sensitivity of bioluminescent detection supports analysis of HiBiT-tagged proteins at endogenous expression levels, helping to avoid artifacts from overexpression. A high-affinity monoclonal antibody specific to HiBiT extends its utility to traditional immunoassay-based applications, offering superior versatility compared to other epitope tags. Here, we demonstrate quantitative capture of HiBiT-tagged targets across a range of endogenous expression levels. Immunoprecipitation with Anti-HiBiT Magne® Beads enabled robust enrichment of tagged proteins and their interaction partners for downstream mass spectrometry analysis. An integrated workflow, combining bioluminescent quantification with antibody-based capture, facilitates the study of endogenous proteins under physiologically relevant conditions