Abstract
Understanding the multiple functions of protein phosphatase 2A (PP2A) rests on elucidating the enzymatic properties of over 50 different possible forms of the PP2A holoenzyme. We describe a procedure for highly purifying each one of these forms. This procedure is based on coexpressing in 293 cells one scaffolding A subunit, one regulatory B subunit, and one catalytic C subunit, each tagged with a different sequence, and purifying the trimeric holoenzyme by three consecutive immunoprecipitations with antibodies against the tags. In a few hours and from a small number of cells, sufficient enzyme can be purified for enzymatic studies. Purification of six different holoenzymes in parallel can easily be accomplished.
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Walter, G., Zhou, J., Ruediger, R. (2007). Purification of PP2A Holoenzymes by Sequential Immunoprecipitation with Anti-Peptide Antibodies. In: Moorhead, G. (eds) Protein Phosphatase Protocols. Methods in Molecular Biology, vol 365. Springer, Totowa, NJ. https://doi.org/10.1385/1-59745-267-X:113
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DOI: https://doi.org/10.1385/1-59745-267-X:113
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-1-58829-711-2
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