Triton X 100 is used to increase the solubility of the protein when used in the lysis buffer for bacterial lysis. But I am not sure how much percentage can be used so that it increases the solubility of my his-tagged protein and doesn't effect the binding of the protein into the Ni-NTA beads. If, someone is using Triton x-100 in their lysis buffer, I would like to know how much final percentage is safe for Ni-NTA protein purification.
The instruction manual for the Cytiva His-trap Ni-affinity column states that the column is compatible with 2% Triton X-100.
https://cdn.cytivalifesciences.com/api/public/content/digi-12993-pdf
Sebastian Schmitt
Thank you for the reply. I am usually following everything u suggested and only 2.5% glycerol. I am trying to use a detergent because the protein I am trying to purify is probably forming aggregates after incubation with the Ni-NTA beads.
Adam B Shapiro Thank you. I will try something ranging from 0.5 % until 2% and hopefully it will do the job.
- Badges
- Science topic