EC Number |
---|
2.4.2.12 | - |
2.4.2.12 | immobilized metal ion affinity chromatography |
2.4.2.12 | immobilized metal ion affinity chromatography (Ni2+) |
2.4.2.12 | lysis of bacteria by French press, elution from Ni-NTA column by 100 and 200 mM imidazole, concentration and desalting, elution from HiLoad Superdex 200GP 26/60 column with 100 mM HEPES pH 7.5, 100 mM NaCl, 10 mM beta-mercaptoethanol, concentrated to 50-75 mg/ml, kept frozen |
2.4.2.12 | lysis of bacteria by ultrasonification in presence of 1 mM phenylmethylsulfonylfluorid (PMSF) and 1 mM dithiothreitol (DTT), ultracentrifugation, supernatant applied to glutathione Sepharose 4B beads and washed, cleavage of glutathione S-transferase (GST) tag by PreScission Protease, purification by gel-filtration chromatography in Tris-HCl pH 8.5, 150 mM NaCl, 1 mM DTT, concentration to 10 mg/ml by ultrafiltration on Microcon YM-10, stored at 4°C |
2.4.2.12 | Ni-NTA resin column chromatography |
2.4.2.12 | partial |
2.4.2.12 | partially frim K-562 leukemic cells |
2.4.2.12 | recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography |