Footprints Without Footwork: Automated Benchtop HRPF for Binding Partners and Interfaces
Move hydroxyl radical protein footprinting onto an Opentrons Flex — and turn a finicky benchtop assay into a reproducible, higher-throughput method.
About this Webinar:
Every protein leaves a footprint — the surface-exposed side chains that are accessible when it folds, when it binds, when it changes shape. Hydroxyl radical protein footprinting (HRPF) reads those footprints by oxidising the exposed residues and leaving the buried ones alone — a structural mass spectrometry technique that runs on standard laboratory equipment plus a benchtop Fenton reaction. Done by hand it is labor-intensive: multiple radical concentrations, replicate reactions, and dose-response curves that are sensitive to pipetting variability. Manual HRPF also limits the technique in scope — applying it across the dozens of pulldowns needed to characterise a complex becomes impractical.
In this webinar we show how moving the workflow onto the Opentrons Flex liquid handler tightens reproducibility and lifts throughput, and how applying that automated workflow to affinity-purified protein complexes lets you identify binding partners and map their interaction interfaces in the same experiment.
What you'll learn in this webinar:
HRPF can be performed using standard laboratory equipment to probe surface-accessible amino acids on proteins and protein complexes
Automating HRPF reactions on the Opentrons Flex reduces variability and increases throughput
Performing HRPF reactions on affinity-purified protein complexes can provide insights into binding partners and interfaces simultaneously
Our Speaker:
Tyler Cropley is a Research Scientist in the Proteomics Laboratory at NYU Langone Health under the supervision of Dr. Beatrix Ueberheide. His work focuses on using mass spectrometry-based methods to study higher-order protein structure. He earned his PhD from Florida State University under the supervision of Dr. Christian Bleiholder.
Register Now