MassIVE MSV000083293

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GFP-fused 53BP1 interacting proteins in HEK293-T cells with and without irradiation

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Description

Experiment: proteins immunoprecipitated with a GFP-fused 53BP1 fragment. Description: GFP-fused 53BP1 fragment was conditionally expressed (Tet-ON) in HEK293-T cells in duplicate. Cells were irradiated (6 Gy) or not, allowed to recover for 24 hours, cross linked with 0.05% paraformaldehyde, lysed with the EZ-ChIP lysis buffer. Lysates were incubated with anti-GFP beads, washed, and eluted by heating in Laemmli buffer. Eluents were run on an SDS-PAGE gel, lanes were cut into four equally sized sections. Sections were separately in-gel digested with trypsin. Non-Irradiated control sample was labeled with O18 by digesting in the presence of O18 water. Duplicates per section of both samples (non-irradiated and irradiated) were combined to give four samples (one per MW range section). Samples were run on the LTQ Orbitrap XL. Data were searched with Mascot (once including both O16 and O18 at the C-terminus, once allowing only O16, once allowing only O18), and analyzed in Scaffold. [doi:10.25345/C5D31N] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: 53BP1 ; irradiation ; DNA damage ; HEK293 ; AP-MS

Contact

Principal Investigators:
(in alphabetical order) 		Laurie Parker, University of Minnesota, USA
Stephen J. Kron, University of Chicago, USA
Submitting User: llparker
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