I tried with Ni-NTA Agarose (Qiagen) using native conditions. I also did Q-Sepharose (ion exchange), followed by Ni-NTA native conditions. Purity was not achieved in either case. Now I'm trying in denatured conditions.
All of your feedback is welcome.
Thank you,
T M P RAJU.
Immobilized Metal Affinity Chromatography (IMAC is used to purify His tagged proteins. Mainly beaded agarose or magnetic particles can be derivatized with chelating groups to immobilize the desired metal ions, which then function as ligands for binding and purification of biomolecules of interest. Expressed His-tagged proteins can be purified and detected easily because the string of histidine residues binds to several types of immobilized metal ions, including nickel, cobalt and copper, under specific buffer conditions. In addition, anti-His-tag antibodies are commercially available for use in assay methods involving His-tagged proteins
