Ideal tools for monitoring protein purification progress
Western blot of His tagged protein probed with Mouse anti Histidine tag antibody, clone AD1.1.10 (MCA1396)
Rabbit anti Mouse IgG:HRP (STAR138A)
Our selection of conjugated anti-His tag antibodies saves you time by removing the need for a secondary antibody.
Like the MBP (maltose binding protein) and GST tag, the His tag is a small epitope tag consisting of between six to nine histidine residues, which is cloned into an expression vector upstream or downstream of a gene of interest. The vector is then introduced into an expression system such as bacteria, yeast or insect cells. The result is the expression of a recombinant protein carrying a polyhistidine tag at its N- or C-terminus.
In order to purify the recombinant protein immobilized metal affinity chromatography (IMAC) is performed. This technique is based on the fact that a cord of histidine residues binds to immobilized metals such as Nickel. As well as His tag antibodies, Bio-Rad offers a variety of kits for the purification of His tagged proteins.
In addition to that anti-histidine antibodies are used to monitor the progress of purification and the fraction containing the recombinant protein by western blotting. Antibodies are also handy in order to ensure no proteolytic degradation of the recombinant protein is taking place.
The IMAC kits are designed for the convenient purification of His-tagged recombinant proteins from cell lysates under native and denaturing conditions. The spin columns ensure a speedy one step process, which removes most contaminants and can achieve purities close to homogeneity. Find out how to do quick protein purifications by reading our IMAC handbook for more information.
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