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Sample preparation of immunoprecipitated EGFP-tagged PARP1 to identify protein-protein interactions

We combined the GFP-Trap® with the fast, sensitive and robust iST sample preparation kit (PreOmics GmbH). The combination of both workflows enables a streamlined sample preparation of immunoprecipitated GFP-fusion proteins for subsequent analysis by mass spectrometry.

In this application note the following process is described:

  • Analyze protein-protein interaction partners of an individual candidate protein in living cells
  • Express EGFP-tagged PARP1
  • Use the GFP-binding protein coupled magnetic beads (GFP-Trap Magnetic Agarose, gtma)
  • Efficiently extract the EGFP-fusion proteins from the cellular extract
  • Combine it with the reproducible iST method