6x-His Tag Monoclonal Antibody (3D5), HRP, Invitrogen™

Description

NOTE: R931-25 was previously sold under product #46-0707. When re-ordering, please use #R931-25. This antibody can be used to detect expression of C-terminal His fusion proteins from bacterial, insect, and mammalian cells. This product contains enough material for 25 Western blots. It is functionally tested using 100 ng of an E. coli expressed fusion protein containing a C-terminal polyhistidine (6xHis) epitope. Chemiluminescent detection should be performed using a 1 minute exposure. For Western blot, dilute in PBS containing 0.05% Tween-20 and 5% nonfat, dry milk (PBSTM). Using chemiluminescence as the detection method, no cross-reactivity has been observed in bacterial lysates. In mammalian lysates, a few cross-reactive proteins have been observed upon overexposure of blots.

The 6x His tag is a synthetic oligo peptide consisting of 6 consecutive histidine residues (HHHHHH). The His tag is commonly expressed as a tag at either N-or C-terminal regions of recombinant proteins to allow isolation or purification by immobilized metal affinity chromatography. Epitope tagging is a technique in which a known epitope is fused to a recombinant protein using genetic engineering. By choosing a particular epitope and recombinant protein combination, epitope tagging makes it possible to detect proteins for which no antibody is available. His epitope tagged proteins that contain a stretch of histidine residues at the carboxyl or amino terminus are used for purification of recombinant proteins by means of metal chelate chromatography due to metal binding properties of histidine clusters on His tagged proteins. For example, the affinity of the histidine-tag motif to Ni2+ by chelation is strong and selective enough to enable purification of the protein to homogeneity by affinity chromatography on a Ni2+-NTA adsorbent. His tag-specific antibodies are used to facilitate detection or coimmunopreciptation of tagged proteins.