Define the interactome of IBTK by Affinity purification coupled with mass spectrometry (AP-MS) and proximity-dependent biotinylation identification (BioID) methods

Affinity purification coupled with mass spectrometry (AP-MS) and proximity-dependent biotinylation identification (BioID) methods are powerful tools to define the interactome for a specific protein bait. Whereas AP-MS results in the identification of proteins that are in a stable complex, BioID labels and identifies proteins that are in close proximity to the bait, resulting in overlapping yet distinct protein identifications. In order to comprehensively characterize the IBTK interactome networks in cells, we developed a tag workflow which allows for both AP-MS and BioID analysis with a single construct, pcDNA5/FRT vector containing FLAG-BirA-IBTK.