Hello.
I wan to evaluate the function of my protein in MTT assay,
I have many problems with the aggregation of this protein after purification. I purify this protein with the IMAC system by GE healthcare device. The elution buffer consists of tris 50 mM, urea 2 M, 2me 5mM, imidazole 300mM, NaCl 500mM. I should preserve this protein in urea 2 molar and 2me 5mM for the reduction of aggregation. For removing of imidazole, I always use dialysis with buffer consists of urea2M, tris 50mM, 2me 5mM, NaCl 150mM.
This protein also can elute by imidazole 200mM. The maximum concentration of this protein in MTT is 40mM. So for MTT, this protein dilutes.
Because in MTT assay I better don't use urea 2 molar and 2me 5mM, can I change the elution buffer to PBS and not remove imidazole?
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