Anti-His-tag (C-terminal); Protein A Purified	back

The availability of larger amounts of proteins by recombinant expression und purification simplifies the analyses for protein structure and function. For purification several strategies have been developed in particular the expression as fusion protein for an efficient application of the affinity chromatography.

In the last couple of years many peptide sequences/epitopes for the purification of recombinant proteins have been established. These so-called “tags” can be used e.g. to determine the cellular localization or to quantify proteins. The polyhistidine “tag” (His-tag) is the most used affinity epitope for the purification of recombinant proteins [1]. Proteins with a polyhistidine tag (e.g.  6xHis or 8xHis) can be purified in one step using a metal-chelate column (e.g. Ni2+, Zn2+, Cu2+ or Co2+) and imidazole as eluent. This method now is a very attractive system for the purification of larger amounts proteins for structural and functional studies. So far His-tagged proteins were successfully purified from different expression systems like E. coli, yeast, insect cells and plant cells [1,2]. An important requirement beside the efficient and robust purification method is the availability of a fast detection system for checking the purification steps of these His-tagged proteins if no specific antibody is available.

In this respect we are pleased to announce that we have developed a very specific and unique rabbit polyclonal antibody against the His-tag [Cat#102-PA80] suitable for detection of His-tagged proteins in the Western analysis and ELISA [3]. The antibody was generated by immunizing rabbits with highly purified His-tagged proteins expressed in insect cells. The anti-His-tag antibodies were then purified from sera by an antigen-affinity chromatography using a His-peptide as matrix. Western analysis with several His-tagged proteins either expressed in insect cells or E. coli showed that the antibody recognizes all tested proteins fused to a C-terminal but not to a N-terminal His-tag. This antibody might be a very good tool to test supernatants or cell lysates for expression of recombinant proteins. For Western analysis the antibody can be used at very low concentration, e.g. at 0.1µg/ml. For more deatils see More Information!

Lit.: 1. Terpe K, Appl. Micrbiol. Biotechnol. 60:523, 2003; 2. Leelavathi et al, Molecular Breeding 11:49, 2003; 3. Bernhardt et al, Laborwelt 5:18, 2004 (German)]

Datasheet (PDF)	back