Quantification of Protein Interaction Network Dynamics using Multiplexed Co-Immunoprecipitation.

ABSTRACT: Dynamic protein-protein interactions control cellular behavior, from motility to DNA replication to signal transduction. However, monitoring dynamic interactions among multiple proteins in a protein interaction network is technically difficult. Here, we present a protocol for Quantitative Multiplex Immunoprecipitation (QMI), which allows quantitative assessment of fold changes in protein interactions based on relative fluorescence measurements of Proteins in Shared Complexes detected by Exposed Surface epitopes (PiSCES). In QMI, protein complexes from cell lysates are immunoprecipitated onto microspheres, and then probed with a labeled antibody for a different protein in order to quantify the abundance of PiSCES. Immunoprecipitation antibodies are conjugated to different MagBead spectral r

SUBMITTER: Brown EA 

PROVIDER: S-EPMC8195492 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature