EC Number   |
|---|
   2.7.1.158 | - |
| 2.7.1.158 | a glutathione affinity column is used to capture intact GST-ZmIPK1. The purified GST-ZmIPK1 fusion is cleaved with thrombin and further purified by a sequential affinity and ion-exchange chromatography. ZmIPK1 protein is eluted at 0.1 M NaCl on a high resolution Mono Q column with an estimated purity of 95%. Identity of the purified protein is confirmed via trypsin digestion followed by matrix-assisted laser deposition/ionization (MALDI) time-of-flight mass spectrometry analysis and comparison of the MALDI fingerprint to the amino acid sequences deduced from ZmIPK1. |
| 2.7.1.158 | by batch-elution from Ni-NTA affinity resin, Analysis by SDS-PAGE gel and Western Blot using an anti-His antibody. Purity of protein is >98% estimated by SDS-PAGE. |
| 2.7.1.158 | expression in Escherichia coli |
| 2.7.1.158 | glutathione agarose bead chromatography |
| 2.7.1.158 | Ni-NTA bead chromatography |
| 2.7.1.158 | recombinant |
| 2.7.1.158 | recombinant enzyme |
| 2.7.1.158 | recombinant His-tagged enzyme from Escherichia coli strain BL21 (Lys) DE3 by nickel affinity chromatography |
| 2.7.1.158 | recombinant LSLt-tagged enzyme from Escherichia coli strain BL21 Star (DE3) by heparin affinity chromatography and gel filtration followed by tag cleavage with TEV protease and again heparin affinity chromatography and gel filtration. The recombinant GST-tagged enzyme from Sf21 insect cells is purified by glutathione affinity chromatography and tag cleavage with PreScission protease, followed by heparin affinity chromatography and gel filtration |
