HIS-Select Affinity Purification

A polyhistidine tag, usually his6, attached to either the amino-terminus or carboxyl-terminus of an expressed recombinant protein enables affinity purification of that protein. Using the principles of immobilized metal affinity chromatography (IMAC), the his-tagged fusion protein binds to a metal chelate-coated substrate and can be eluted with imidazole. In addition to plant-plant proteomic applications, His-tagged proteins from bacterial or animal sources may be expressed in plants to avoid cross-contamination of the purified protein with other bacterial or animal proteins.1 Our HIS-Select nickel affinity system uses tetradentate chelated nickel for high selectivity bound to its support with a neutral spacer to minimize unwanted ionic interactions. HIS-select nickel affinity gel can bind about 15 mg protein/ml. The ligand has the same high capacity on highly crosslinked agarose for fast flow and high pressure chromatography, and in the EZview Red agarose format. HIS-select spin columns are good for one-step purification of his-tagged proteins from small batches of crude extract. Finally, the nickel-chelate complex bound to 96 well plates enables high-throughput screening for quantification or preparation for MALDI mass spectrometry.

1. MacRae, A.F., Preiszner, J., NG, S., and Bolla, R.I. Expression of His-tagged Shigella IpaC in Arabidopsis. J. Biotechnol 112:247. 2004

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E3528 EZview Red HIS-Select® HC Nickel Affinity Gel
H8162 HIS-Select® Cobalt Affinity Gel
H0537 HIS-Select® HF Nickel Affinity Gel
P6611 HIS-Select® Nickel Affinity Gel
H7787 HIS-Select® Spin Columns