EC Number   |
|---|
   3.1.3.67 | - |
| 3.1.3.67 | fusion-protein is purified by Glutathione-Sepharose-4B affinity chromatography |
| 3.1.3.67 | Poly-His tag proteins are purified with a HisTrap HP kit using buffers with 10 mM mercaptoethanol, enzymes are further purified with a gel filtration column in 100 mM NaCl, 10 mM Tris, pH 7.4, and 1 mM dithiothreitol. Final purification is done with a anion exchange column in 10 mM Tris, pH 7.4 with a linear gradient from 50-600 mM NaCl. |
| 3.1.3.67 | recombinant enzyme |
| 3.1.3.67 | recombinant PTEN is purified to near homogeneity using four sequential column chromatographic steps: a diethylaminoethyl (DEAE) Sepharose anion exchange column, a bio-gel hydroxyapatite HT (HAP) column, a Mono-S cation exchange column, and a Mono-Q anion exchange column |
| 3.1.3.67 | treatment with alkaline phosphatase fully dephosphorylates the phosphorylation sites. Unphosphorylated PTEN and alkaline phosphatase can be separated by ion exchange column chromatography |
