His-tag His-tagged protein is effectively retained on

His-tag provides a way to specifically detect or
purify proteins of interest without the need to use protein-specific antibody
or probes (Waugh, 2011). Indeed
His- tag has been widely used to enhance protein purification by several
techniques one of which is called immobilized metal affinity chromatography (Randolph, 2011). The underlying basis of IMAC method is the
interaction between a
transition metal ion such as copper (Cu2+), cobalt (Co2+), nickel
(Ni2+), zinc (Zn2+), calcium (Ca2+), and Iron III (Fe3+) immobilized on a matrix and specific amino acid side
chains (Randolph, 2011). Histidine is one of the amino acids that can
strongly interact and form coordinate bonds with the immobilized transition
metal ions (Bang and Kent,
2005). This allows purification of  His-tagged protein from complex mixtures that
is only His-tagged protein is effectively retained on IMAC column matrices (Bang and Kent,
2005). The
type of metal ion used in IMAC is based on the objective of the application.

For example, if the primary concern was His-tagged protein purification then
cobalt is the ideal ion metal to use since it exhibits more specific
interaction with His-tag than all of the other metal ions (Bang and Kent,
2005). His-tagged protein can be easily eluted from the column by either
adjusting the buffer pH or adding free imidazole to the column buffer

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