AI Article Synopsis
- Immunoprecipitation has been the traditional method for isolating native protein complexes, but it relies heavily on antibody quality and consumes these antibodies, which limits further analyses.
- An alternative method using affinity purification is introduced, utilizing human Flp-In cells that express a Protein A-tagged version of the PEX5L receptor.
- This new method allows for the effective isolation of both soluble and membrane-bound complexes containing PEX5L through an established affinity-based approach.
Article Abstract
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.
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| http://dx.doi.org/10.1007/978-1-0716-3048-8_26 | DOI Listing |
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