Updated project metadata.
Identification of interacting partners of the Partner and Localizer of BRCA2 (PALB2), essential regulator of DNA repair by homologous recombination. Interacting partners of PALB2 were purified by GFP pull down from asynchronous HEK293 Flp-In T-REx cells, exogenously expressing Flag-EGFP tagged PALB2 at a similar level than the endogenous PALB2 level. Before GFP pull down, whole cell protein extracts were pre-cleared on IgG agarose beads to decrease binding of non-specific proteins during GFP pull down. GFP pull down were performed using GFP-Trap agarose beads (Chromotek) and washed at low salt concentration (150mM NaCl), to maintain interactions of low affinity binding protein partners. As a negative control, Flag-EGFP interacting proteins were purified, following the same protocol as described for the purification of Flag-EGFP PALB2 interacting proteins. Experiments were performed in triplicate.