Abstract
A polyhistidine-tag is an amino acid motif that contains at least six histidine (His) residues, usually at the N- or C-terminus of the protein. This tag can also be referred to as a hexa histidine-tag or a 6x His-tag. The protocol described here has been developed to purify His-tagged proteins from E. coli under denaturing conditions using Ni-NTA agarose beads.
Materials and Reagents
Equipment
Procedure
Notes
The amount of culture required for an experiment like this will depend on the level at which the protein is expressed, which must be determined empirically for each expression experiment.
Recipes
Acknowledgments
This protocol has been modified and adapted in the Espenshade Lab, Johns Hopkins School of Medicine. Funding to support different projects that have used this protocol has come from NIH – National Heart, Lung, and Blood Institute, National Institute of Allergy and Infectious Diseases, the Pancreatic Cancer Action Network, and the American Heart Association.
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.
Hi Alexander,Thank you for pointing out. With author's permission, we have corrected it in the text accordingly.--Bio-protocol Editorial Board