This will not work. Ni metal (s) will not interact with high affinity on a His patch (his-tag). Nickel salts would in several cases (NiCl) but it is not magnetic
On Tuesday, January 17, 2017 at 9:50:53 AM UTC-5, CoreyHowe wrote:
-- On Tuesday, January 17, 2017 at 9:50:53 AM UTC-5, CoreyHowe wrote:
A friend and I were talking about a simplified His-tag purification protocol for a peptide we'd like to produce and use, and I wanted to see what other people thought of it's feasibility.Instead of applying the cell lysate to a nickel column, first add nickel powder ($5 on ebay) to the lysate in a tube and mix, once properly mixed then apply a strong magnet to the outside of the tube to attract the nickel bound to the his-tag, pour off the excess lysate, lower the pH with an acid to elute the peptide from the nickel, and finally transfer to a new tube.Could this be feasible?
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