PTEN is a one of the most frequently mutated tumor suppressors in human cancers. It is essential for regulating diverse biological processes and through its lipid phosphatase activity regulates the PI 3-Kinase signaling pathway. Sensitive phosphatase assays are employed to study the catalytic activity of PTEN against phospholipid substrates. Here we describe protocols to assay PTEN lipid phosphatase activity using either purified enzyme (purified PTEN lipid phosphatase assay) or PTEN immunopurified from tissues or cultured cells (cellular IP PTEN lipid phosphatase assay) against vesicles containing radiolabeled PIP3 substrate.
|Title of host publication||Protein Tyrosine Phosphatases|
|Number of pages||11|
|Publication status||Published - 2016|
|Name||Methods in Molecular Biology|
Spinelli, L., & Leslie, N. R. (2016). Assays to Measure PTEN Lipid Phosphatase Activity In Vitro from Purified Enzyme or Immunoprecipitates. In Protein Tyrosine Phosphatases (pp. 95-105). (Methods in Molecular Biology; Vol. 1447). Springer. https://doi.org/10.1007/978-1-4939-3746-2_6