Hello Everyone, We included 10 mM EDTA in our Ni-NTA and gel filtration buffers to prevent aggregation of our protein, as suggested by the literature (indeed, excluding EDTA led to precipitation during the concentration steps). In the presence of 10 mM EDTA, we were able to concentrate the protein to >50 mg/mL without seeing any precipitation. We would now like to carry our protein forward into crystallization trials. I am just wondering if anyone observed adverse effects when screening for crystals in EDTA-containing buffer? Is there any reason for us to try to reduce/dilute the concentration of EDTA? Our enzyme is not metal-dependent.
Thanks in advance, Gyl
