Affinity Purification of Soluble and Membrane-Bound Protein Complexes by a FlpIn Strategy.

For a long time, the isolation of native protein complexes from human cells was accomplished by immunoprecipitation experiments. However, success depends on the quality of the antibodies and the method consumes valuable antibodies, which can hinder subsequent analysis of the isolated complexes. Here, we demonstrate an alternative approach based on affinity purification. It utilizes human Flp-In cells, which genomically express a Protein A-tagged version of the human peroxisomal import receptor PEX5L. Native soluble and membrane-bound complexes containing PEX5L can thereby be isolated via a well-known affinity-based strategy.