Ethylenediaminetetraacetic Acid → Imidazole-4-acetic acid

Participants

Reaction Steps

The cell pellet was homogenized in 6 volumes 0.1 M Tris-HCl pH 8, 0.2 M NaCl, 5 mM DTT, 1 mM EDTA, and Apo-2L was isolated from the soluble fraction by IMAC on a chelating hiTRAP column (Pharmacia) charged with nickel. The Apo-2L had a weak affinity for immobilized metal and could be eluted with low concentrations of imidazole. A final purification was obtained by cation exchange chromatography on a SP hiTRAP column (Pharmacia). Concentrations of purified Apo-2L variants were determined by absorbance measurements using an e280 of 1.4 mg−1 ml cm−1.

Provenance & Evidence